Minimally Manipulated Bone Marrow-Derived Cells Can Be Used for Tissue Engineering In Situ and Simultaneous Formation of Personalized Tissue Models

Abstract

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 tissueengineered 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.

Adverse events, side effects and complications in mesenchymal stromal cell-based therapies

“…MSCs were recognised as easy derivable and the most applicable choice among the other cell-sources for a wide range of pathologies (5)(6)(7)(8)(9)(10). These cells are commonly obtained from the bone marrow (11), adipose tissue (12), umbilical cord (13), dental pulp (14), gingiva (15), perinatal tissues (16), etc.…”

Intraoperative Creation of Tissue-Engineered Grafts with Minimally Manipulated Cells: New Concept of Bone Tissue Engineering In Situ

“…If adhesion is achieved, the odds of cell persistence in the transplantation area are increased. Previously, we showed that 20 min incubation in cell suspension was enough for the adhesion of BM-MNCs on bone material [ 20 ]. In another study, fluorescence microscopy confirmed that rat BM-MNCs adhered to different types of scaffolds after 10 min of incubation at 37 °C [ 65 ].…”

“…Multiple clinical and animal studies demonstrated results in improved bone healing [ 4 , 18 , 19 ]. Importantly, BM-MNCs are available as minimally-manipulated cells without preliminary cell culture [ 20 ]. BM-MNCs are a heterogeneous population of cells with a single nucleus, which includes mesenchymal stromal and stem cells, hematopoietic stem cells, endothelial progenitor cells, very small embryonic-like stem cells, embryonic stem cells, etc.…”

“…Despite numerous types of scaffolds, bone graft autotransplantation is still considered a ‘gold-standard’ approach to repairing bone defects ( Figure 2 a). However, a well-known method of autologous osteoplasty is commonly limited by the harvested bone volume and associated with an additional risk of complications in the donor site [ 20 , 54 ].…”