Abstract
Autologous bone transplantation is the principal method for reconstruction of large bone defects. This technique has limitations, such as donor site availability, amount of bone needed and morbidity. An alternative to this technique is tissue engineering with bone marrow-derived mesenchymal stem cells (BMSCs). In this study, our aim was to elucidate the benefits of culturing BMSCs in 3D compared with the traditional 2D culture. In an initial screening, we combined BMSCs with four different biogels: unmodified type I collagen (Col I), type I collagen methacrylate (ColMa), an alginate and cellulose-based bioink (CELLINK) and a gelatin-based bioink containing xanthan gum (GelXA-bone). Col I was the best for structural integrity and maintenance of cell morphology. Osteogenic, adipogenic, and chondrogenic differentiations of the BMSCs in 2D versus 3D type I collagen gels were investigated. While the traditional pellet culture for chondrogenesis was superior to our tested 3D culture, Col I hydrogels (i.e., 3D) favored adipogenic and osteogenic differentiation. Further focus of this study on osteogenesis were conducted by comparing 2D and 3D differentiated BMSCs with Osteoimage® (stains hydroxyapatite), von Kossa (stains anionic portion of phosphates, carbonates, and other salts) and Alizarin Red (stains Ca2+ deposits). Multivariate gene analysis with various covariates showed low variability among donors, successful osteogenic differentiation, and the identification of one gene (matrix metallopeptidase 13, MMP13) significantly differentially expressed in 2D vs. 3D cultures. MMP13 protein expression was confirmed with immunohistochemistry. In conclusion, this study shows evidence for the suitability of type I collagen gels for 3D osteogenic differentiation of BMSCs, which might improve the production of tissue-engineered constructs for treatment of bone defects.
Highlights
Despite the innate capabilities of physiological regeneration of bone tissue, a current challenge in the fields of orthopedics and odontology is the reconstruction of large bone tissue defects due to trauma, and congenital and other pathological conditions, such as cancer [1,2].The current gold standard for bone reconstruction is autologous bone grafting with or without attached vascularization
We evaluated various commercially available biogels that can be combined with bone marrow-derived mesenchymal stem cells (BMSCs) for osteogenic differentiation
The BMSC cultures were analyzed by flow cytometry for various stem cell surface markers
Summary
Despite the innate capabilities of physiological regeneration of bone tissue, a current challenge in the fields of orthopedics and odontology is the reconstruction of large bone tissue defects due to trauma, and congenital and other pathological conditions, such as cancer [1,2]. The current gold standard for bone reconstruction is autologous bone grafting with or without attached vascularization. This approach, albeit effective, is associated with elevated donor-site infection, acute pain, and morbidity [3,4]. The field has further advanced by utilizing novel therapies with biomaterial scaffolding, in vitro creation of cellular constructs and use of decellularized bone for scaffolding [6,7]
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