Osteogenic extracellular matrix sheet for bone tissue regeneration

The application of extracellular matrix (ECM) sheets without a scaffold is not extensively reported in bone regenerative medicine. The aim of the present study was to demonstrate that an osteogenic ECM sheet (OECMS) can retain ECM integrity and growth factors to enhance bone formation in a rat non-union model. OECMS was produced from osteogenic cell sheets (OCS). Collagen and growth factor [bone morphogenetic protein 2 (BMP-2), vascular endothelial growth factors (VFGFs), basic fibroblast growth factor (bFGF) and transforming growth factor β1 (TGF-β1)] concentrations in the OECMS were quantified by enzyme-linked immunosorbent assay (ELISA). Next, hydroxyapatite (HA) constructs combined with OECMSs were implanted subcutaneously into the rats' backs to evaluate their osteoinductive capacity by histological evaluation. In addition, OECMSs were implanted in a rat femoral non-union model. 18 male Fischer 344 inbred rats were divided into OECMS and control groups. Fracture healing was evaluated by radiological and histological analyses at 2, 5 and 8 weeks and biological analysis at 8 weeks. Collagen I and growth factors were retained in the OECMSs. Osteoid formation was identified in the HA combined with OECMS at 4 weeks. Enhanced bone regeneration at the non-union of the OECMS group was confirmed at 5 and 8 weeks. Biomechanical testing revealed a significantly higher maximum bending load in the OECMS group as compared to the control group at 8 weeks. The results demonstrated that OECMS retained BMP-2 and TGF-β1 and high osteoinductive and osteoconductive capacity. As such, OECMS represents a potential new scaffold-free material for bone tissue engineering

A practical and effective strategy in East Asia to prevent anti‐D alloimmunization in patients by C/c phenotyping of serologic RhD‐negative blood donors

Abstract Serologic RhD‐negative red cells can cause anti‐D alloimmunization if they carry the Asian‐type DEL or other DEL variants. RHD genotyping is a viable countermeasure if available, but inexpensive alternatives are worthy of consideration. RhD‐negative blood donors in Japan were studied by anti‐D adsorption‐elution and RHD genotyping. We collated published case reports of RhD‐negative red cell transfusions associated with inexplicable anti‐D immunization. Of 2754 serologic RhD‐negative donors, 378 were genotyped D/d. Anti‐D adsorption‐elution revealed 63.5% (240 of 378) to be DEL, of whom 96.7% (232 of 240) had the 1227G > A variant, diagnostic for the Asian‐type DEL. All 240 donors also carried at least one C antigen; none had a cc phenotype. The chance of transfusing DEL red cells to genuinely RhD‐negative Asian patients (based on a three‐unit transfusion) ranges from 16.7% in Korea to 69.4% in Taiwan, versus 0.6% in Germany. Among 22 RhD‐negative recipients of serologic RhD‐negative red cells, who produced new or increased anti‐D antibody titers, all 17 from East Asia were transfused with red cells with a C‐positive phenotype or known to be Asian‐type DEL or both. Serologic RhD‐negative East Asians with a cc phenotype can be red cell donors for RhD‐negative recipients, especially those of childbearing potential