Objective To evaluate the efficiency of our self-designed enriched filtrator of stem cells in screening and collecting enriched human bone marrow stem cells (BMSCs) for rapid construction of bioactive materials.Methods Bone marrow blood from 32 patients was circulated in our self-designed enriched filtratorand filtered through beta-tricalcium phosphate(β-TCP).The numbers of BMSCs were counted pre-and post-filtration to assess the enrichment efficiency.The numbers of nucleated cells,red blood cells and platelets were compared pre-and post-filtration to assess the effects of the enriched filtrator of stem cells on blood cells.Bacteriological examinations were performed to test the safety of the device.Scanning electron microscopy was carried out to observe the adhesion of BMSCs on β-TCP.Results There was a significant difference between the pre-fihration colony number of BMSCs cultured (232.8 ± 128.6/mL) and the post-filtration one (37.8 ± 34.9/mL) (t =10.743,P ＜ 0.001).The efficiency of BMSCs enrichment of the device was 85.6％ ±8.4％ (from 63.8％ to 97.1％).There were no significant differences between pre-filtration and post-filtration in the numbers of nucleated cells [(16.8 ± 5.4) × 109/mL versus (16.3 ±5.0) ×l09/mL],red blood cells [(3.9±0.6) ×1012/mL versus (3.8±0.6) ×1012/mL] and platelets [(120.4 ± 54.6) × 109/mL versus (123.0 ± 51.8) × 109/mL] (P ＞ 0.05).All the bacteriological examinations of the bone marrow samples were negative.After in vitro osteogenic induction culture,formation of numerous BMSCs colonies was observed on the β-TCP by scanning electron microscopy.Conclusion Since our self-designed enriched filtrator can efficiently collect enriched BMSCs and combine them with β-TCP simultaneously,the device is capable of rapid and safe construction of bioactive materials.

Objective To evaluate the validity and security of our self-designed bone marrow stem cell Screen-Enrich-Combine(-Biomaterials) Circulating System(SECCS) for bone non-union of limbs. Methods From May 2013 to December 2015, 24 patients with limb non-union were treated at our de-partment. They were aged from 20 to 65 years (mean, 42.8 years). Their non-union involved femur in 17 cases, tibia in 4, radius in one, humerus in one and fibula in one. In surgery, 80 mL bone marrow blood was aspirated from the anterior superior iliac spine for rapid preparation of bone substitute(β-TCP)composite with bone marrow stem cells by SECCS which was then implanted at the non-union locations. The bone marrow blood was collected before and after enrichment for stem cell counts. The bone union, clinical union time and related com-plications were evaluated by follow-up X-rays at 1, 3, 6 and 9 months after surgery. Results Each collection of bone marrow blood took 13.5 minutes on average. The enrichment rate of stem cells was 81.2%. 11,751 ± 1,011 stem cells were implanted per patient on average. All the patients were followed up for 9 to 48 months (mean, 19.2 months). Twenty-two patients acquired clinical union 9 months after operation and the other 2 suffered from malunion due to insufficient bone implant, yielding a union rate of 91.6% and an average union time of 6.5 months. No graft infection or internal fixation failure occurred and no severe complications hap-pened at the donor or implant sites.Conclusion The bioactive bone substitute manufactured by our self-designed SECCS can be used as a novel therapy for limb non-union, and this device is moreover charac-terized with convenience, limited invasion and satisfactory osteogenesis so that complications of autologous bone transplantation can be avoided.

Objective To introduce a new method for preparation of bioactive β-tricalcium phosphate (β-TCP) by rapid stem cell screen-and-enrich-and-combine circulating system (SECCS) and evaluate its efficacy in the treatment of fresh fractures and bone defects.Methods Twenty-two patients with fresh fracture and bone defects were treated with SECCS from July 2013 to April 2016.They were 16 males and 6 females with an average age of 52.2 years (from 27 to 81 years).There were 15 tibial plateau fractures and 7 calcaneal fractures.The average size of bone defects was 12.5 mL.Bioactive β-TCP was prepared by SECCS intraoperatively and implanted back immediately into the bone defects.Radiographic examination,Lysholm knee scoring and Maryland foot scoring were used for assessment of curative efficacy.Results The 22 patients were followed up for an average of 25.7 months (from 12 to 46 months).By SECCS,the enrichment efficiency of bone marrow stromal cells (BMSCs) reached up to 82.4％ and the cell viability was not affected.The tibial plateau fractures were re-transplanted with 13,381.3 BMSCs on average and healed after an average of 8.9 weeks (from 6 to 15 weeks).The Lysholm knee scores at one year postoperatively averaged 93.6 points (from 84 to 100 points),yielding 10 excellent cases,4 good cases and one fair case.The calcaneal fractures were implanted back with 16,677.7 BMSCs on average and healed after an average of 9.4 weeks (from 8 to 13 weeks).The average Maryland foot score at one year after operation was 93.6 points (from 85 to 98 points),yielding 6 excellent cases and one good case.Conclusion Bioactive materials prepared by SECCS are good bone grafts for fresh fractures and bone defects.