Intraoperative enrichment of bone autograft with autologous bone marrow-derived mononuclear cells isolated simultaneously: moving towards tissue engineering in situ

Background: The use of tissue-engineered bone autografts is a promising approach for bone defects restoration. The isolation of cells and their seeding on bone autograft is usually carried out in a laboratory, requiring significant time and two separate surgical interventions. Intraoperative creation of tissue-engineered bone autograft can represent a perspective solution. The aim of this study is to investigate the possibility of creation of tissue-engineered bone autograft by intraoperative enrichment of bone tissue with bone marrow-derived mononuclear cells (BM-MNCs) isolated simultaneously. Methods: Red bone marrow and autologous bone tissue (bone fragments and bone chips) of the donor were harvested intraoperatively. BM-MNCs were isolated, and bone fragments were enriched with BM-MNCs intraoperatively. Assessment of the adhesion and proliferation of BM-MNCs on bone fragments was carried out by fluorescence microscopy and histological examination. MTT assay was used to compare metabolic activity of BM-MNCs and wBMA cells seeded on bone chips. Results: Autologous bone fragments were colonized with autologous BM-MNCs isolated simultaneously in the O.R. with further adhesion and active growth of cells. When seeded on bone chips, metabolic activity of BM-MNCs was statistically significantly higher compared to wBMA cells (p-value=0.0272) on day 14. There was no difference in metabolic activity of BM-MNCs and wBMA cells cultured in nutrient medium without bone chips. Conclusion: Technically simple method of intraoperative enrichment of autologous bone fragments with BM-MNCs isolated simultaneously allowed to create tissue-engineered bone autograft in the O.R. The safety and effectiveness of intraoperatively enriched autografts should be investigated further.