Cancellous bone allograft seeded with human mesenchymal stromal cells: a potential good manufacturing practice-grade tool for the regeneration of bone defects |
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| Authors: | Maik Stiehler F. Philipp Seib Juliane Rauh Anja Goedecke Carsten Werner Martin Bornhäuser Klaus-Peter Günther Peter Bernstein |
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| Affiliation: | 1. Department of Orthopaedics, University Hospital Carl Gustav Carus, Dresden, Germany;2. Center for Regenerative Therapies, Dresden, Germany;3. Leibniz Institute for Polymer Research and Max Bergmann Center for Biomaterials, Dresden, Germany;4. Medical Clinic I, University Hospital Carl Gustav Carus, Dresden, Germany;1. Associate Professor, College of Dentistry, The University of Iowa, Iowa City, IA;1. Dentistry Faculty, Oral and Maxillofacial Department, Pamukkale University, Denizli, Turkey;2. Dentistry Faculty, Oral and Maxillofacial Department, Cumhuriyet University, Sivas, Turkey;3. Medicine Faculty, Pathology Department, Necmettin Erbakan University, Konya, Turkey;1. Department of Oral and Maxillofacial Surgery, Hannover Medical School, Germany;2. Department of Small Cloven-Hoofed Animals, University of Veterinary Medicine Hannover, Germany;3. Department of Small Animals, University of Veterinary Medicine Hannover, Germany;4. Department of Traumatology and Regenerative Joint Surgery, Hannover Medical School, Germany;1. Assistant Professor, Department of Oral and Craniomaxillofacial Surgery, University Medical Center Freiburg, Freiburg, Germany;2. Resident, Department of Oral and Craniomaxillofacial Surgery, University Medical Center Freiburg, Freiburg, Germany;3. Professor, Department of Oral and Craniomaxillofacial Surgery, University Medical Center Freiburg, Freiburg, Germany; and Department of Oral and Maxillofacial Surgery, University of São Paulo Faculty of Dentistry, Ribeirão Preto, São Paulo, Brazil;4. Professor, Department of Oral and Craniomaxillofacial Surgery, University Medical Center Freiburg, Freiburg, Germany;6. Associate Professor, Department of Oral and Craniomaxillofacial Surgery, University Medical Center Freiburg, Freiburg, Germany |
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| Abstract: | Background aimsCombining autologous bone precursor cells with cancellous bone allograft (CBA) offers an appealing strategy for skeletal regeneration. In this context, multipotent mesenchymal stromal cells (MSC) provide an excellent cell source because they are readily harvested from donors, expanded and differentiated in vitro. The aim of this study was to evaluate the proliferation, morphology, osteogenic differentiation and stem cell-related gene expression during static long-term ex vivo cultivation using human MSC and CBA under good manufacturing practice (GMP)-conforming conditions.MethodsMSC were isolated from healthy donors (n = 5) and cultivated on peracetic acid-sterilized CBA in the presence of 10% human platelet-rich plasma without osteogenic supplements. Total protein content, cell-specific alkaline phosphatase (ALP) activity and osteogenic marker gene expression levels were assessed. Stem cell-related gene expression was compared with MSC monolayer cultivation using microarray analysis. Furthermore, cellular distribution and morphology within the porous CBA were visualized by histology and scanning electron microscopy.ResultsEffective adhesion, spreading, proliferation and intercellular contact of human MSC within the pores of CBA were observed during the study (≤42 days). Cell-specific ALP activity peaked after 3 weeks of cultivation. Gene expression of early, intermediate and late osteogenic marker genes was detectable during long-term cultivation. Microarray-based annotation and biologic interaction network data analysis indicated that expression levels of genes encoding crucial differentiation-regulating proteins and extracellular matrix components involved in the process of osteogenesis were induced in CBA-cultivated MSC.ConclusionsMSC-vitalized CBA offers an attractive GMP-grade bone-filling material. Further research is warranted to evaluate its bone-healing potential in vivo. |
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