Pierini Michela, Lucarelli Enrico, Duchi Serena, Prosperi Susanna, Preve Eleonora, Piccinini Marzio, Bucciotti Francesco, Donati Davide
Osteoarticular Regeneration Laboratory, 3rd Orthopaedic and Traumatologic Division, Rizzoli Orthopaedic Institute, Bologna, 40136, Italy.
Department of Biomedical and Neuromotor Sciences (DIBINEM), Alma Mater Studiorum University of Bologna, Bologna, 40123, Italy.
J Biomed Mater Res B Appl Biomater. 2016 Jul;104(5):894-902. doi: 10.1002/jbm.b.33441. Epub 2015 May 7.
The purpose of this study was to examine the in vitro cytocompatibility of a novel injectable multiphasic bone substitute (MBS) based on polysaccharide gel-coated OSPROLIFE(®) hydroxyapatite (HA)/tetracalcium phosphate (TTCP) granules combined with bone marrow concentrate (BMC). Polysaccharide gel-coated granules loaded in syringe were combined with BMC diluted in ionic crosslinking solution. The product was then maintained in culture to investigate the cytocompatibility, distribution, and osteogenic differentiation function of cells contained in the BMC. The in vitro cytocompatibility was assessed after 0, 24, and 96 h from the injectable MBS preparation using the LIVE/DEAD(®) staining kit. The results highlighted that cells remained viable after combination with the polysaccharide gel-coated granules; also, viability was maintained over time. The distribution of the cells in the product, observed using confocal microscopy, showed viable cells immersed in the polysaccharide gel formed between the granules after ionic crosslinking. The mesenchymal stromal cells (MSC) contained in the injectable MBS, the basic elements for bone tissue regeneration, were able to differentiate toward osteoblasts, producing an osteogenic matrix as evidenced by alizarin red-s (AR-S) staining. In conclusion, we found that the injectable MBS may have the potential to be used as a bone substitute by applying a "one-step" procedure in bone tissue engineering applications. © 2015 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater, 104B: 894-902, 2016.
本研究的目的是检测一种新型可注射多相骨替代物（MBS）的体外细胞相容性。该骨替代物基于多糖凝胶包被的OSPROLIFE®羟基磷灰石（HA）/磷酸四钙（TTCP）颗粒，并与骨髓浓缩物（BMC）相结合。将装在注射器中的多糖凝胶包被颗粒与在离子交联溶液中稀释的BMC混合。然后将该产物置于培养环境中，以研究BMC中所含细胞的细胞相容性、分布及成骨分化功能。使用LIVE/DEAD®染色试剂盒，在可注射MBS制备后的0、24和96小时评估其体外细胞相容性。结果表明，细胞与多糖凝胶包被颗粒混合后仍保持活力，并且随着时间推移活力得以维持。通过共聚焦显微镜观察产物中细胞的分布，结果显示活细胞浸没在离子交联后颗粒间形成的多糖凝胶中。可注射MBS中含有的间充质基质细胞（MSC）是骨组织再生的基本要素，能够向成骨细胞分化，产生成骨基质，茜素红S（AR-S）染色证明了这一点。总之，我们发现这种可注射MBS在骨组织工程应用中采用“一步法”时，可能有潜力用作骨替代物。© 2015威利期刊公司。《生物医学材料研究杂志》B部分：应用生物材料，104B: 894 - 902，2016年。
