Abstract
BackgroundWhen bone marrow is repeatedly filtered through porous material, the mesenchymal stem cells (MSCs) in the bone marrow can adhere to the outer and inner walls of the carrier material to become enriched locally, and this is a promising method for MSC enrichment. In this process, the enrichment efficiency of MSCs involved in the regulation of the cell ecology of postfiltration composites containing other bone marrow components is affected by many factors. This study compared the enrichment efficiency and characterized the phenotypes of enriched MSCs obtained by the filtration of autologous bone marrow through different porous bone substitutes.MethodsHuman bone marrow was filtered through representative porous materials, and different factors affecting MSC enrichment efficiency were evaluated. The soluble proteins and MSC phenotypes in the bone marrow before and after filtration were also compared.ResultsThe enrichment efficiency of the MSCs found in gelatin sponges was 96.1% ± 3.4%, which was higher than that of MSCs found in allogeneic bone (72.5% ± 7.6%) and porous β-TCP particles (61.4% ± 5.4%). A filtration frequency of 5–6 and a bone marrow/material volume ratio of 2 achieved the best enrichment efficiency for MSCs. A high-throughput antibody microarray indicated that the soluble proteins were mostly filtered out and remained in the flow through fluid, whereas a small number of proteins were abundantly (> 50%) enriched in the biomaterial. In terms of the phenotypic characteristics of the MSCs, including the cell aspect ratio, osteogenetic fate, specific antigens, gene expression profile, cell cycle stage, and apoptosis rate, no significant changes were found before or after filtration.ConclusionWhen autologous bone marrow is rapidly filtered through porous bone substitutes, the optimal enrichment efficiency of MSCs can be attained by the rational selection of the type of carrier material, the bone marrow/carrier material volume ratio, and the filtration frequency. The enrichment of bone marrow MSCs occurs during filtration, during which the soluble proteins in the bone marrow are also absorbed to a certain extent. This filtration enrichment technique does not affect the phenotype of the MSCs and thus may provide a safe alternative method for MSC enrichment.
Highlights
When bone marrow is repeatedly filtered through porous material, the mesenchymal stem cells (MSCs) in the bone marrow can adhere to the outer and inner walls of the carrier material to become enriched locally, and this is a promising method for MSC enrichment
After bone marrow filtration at the aforementioned three ratios, the enrichment efficiency of the bone marrow MSCs was 71.2% ± 8.9%, 55.6% ± 14.2% and 40.8% ± 15.9%, respectively; the efficiency was significantly higher at a ratio of 2:1 than at a ratio of 7:1 (P = 0.032)
MSC enrichment technology, especially filtration enrichment, can lead to the direct adhesion of MSCs to the Specific factors influencing the enrichment efficiency of MSCs When using the enrichment technique to prepare bioactive materials rich in MSCs for bone repair, a surgeon first selects bone substitutes of the appropriate type and volume according to the preoperative evaluation results, collects a certain volume of bone marrow to be filtered through the material multiple times, and implants the prepared composite material into the bone repair region
Summary
When bone marrow is repeatedly filtered through porous material, the mesenchymal stem cells (MSCs) in the bone marrow can adhere to the outer and inner walls of the carrier material to become enriched locally, and this is a promising method for MSC enrichment. In this process, the enrichment efficiency of MSCs involved in the regulation of the cell ecology of postfiltration composites containing other bone marrow components is affected by many factors. The bone repair effect is reliable, the approach used to obtain sufficient seed cells by in vitro culturing is accompanied by several drawbacks, including long culturing times, expensive procedures and equipment, high costs, and the ethical controversy related to the use of animal serum, all of which make this approach difficult to implement in clinical practice
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