Collagen is the main component of the extra-cellular matrix and has been utilised for numerous clinical applications in many forms and products. However, since collagen remains a natural animal-derived biopolymer, variation between batches should be addressed and minimised to ensure reproducibility of the fabrication process. Recently, electro-spinning of collagen has been introduced as a leading technique for the production of bio-mimetic nano-scale scaffolds for tissue-engineering applications. However, no protocols are available that would allow comparisons of the quality of different collagen raw materials prior to the electro-spinning process. In order to bridge this gap we assessed the solubility of various freeze-dried collagens in 0.5 M acetic acid and analysed the solved collagen by gel electrophoresis. We show that raw material of limited solubility in acetic acid will not render high quality electro-spun nano-fibres using hexafluoropropanol. In particular, insoluble collagen directly failed to produce nano-fibres, collagen of reduced solubility produced fused nano-fibres with limited inter-nano-fibre space, whilst purified type-I collagen of high solubility produced smooth, reproducible nano-fibres. Gel electrophoresis confirmed the amount of solubility, as well as qualitative differences in terms of collagen cross-links and collagen types. We recommend this simple and fast step to save costs and to enhance control over the electro-spinning process of collagen. Furthermore, we believe that the solubility test should be introduced prior to any collagenous matrix preparation in order to ensure reproducibility and accuracy.
661 bp coding for the carboxyl end of the large sub-unit of EBV ribonucleotide reductase was cloned into the pMal plasmid vector. Purified recombinant protein was tested in IgG and IgA ELISAs. For the IgG assay, 81 out of 100 NPC patients tested positive, whereas for the IgA assay, 60 tested positive. Among 100 normal individuals, I tested positive for the IgG assay and 9 tested positive for the IgA assay. The IgG assay picked up 6 out of 19 NPC sera which were IFA-VCA- and IFA-EA-negative for IgA antibodies. Hence the recombinant ribonucleotide reductase could have good potential as a diagnostic test for NPC or could serve as a complementary test to IFA.
Acute graft-versus-host disease (GVHD) is still a major hurdle for successful bone marrow transplantation (BMT). Although many immunosuppressive drugs are available, none of them alone or in combination are able to completely abolish acute GVHD. The lifelong immunosuppression profoundly reduces the quality of life of BMT recipients. Therefore, new therapeutic approaches are needed. We previously reported that, in an acute GVHD model using SCID mice as recipient, incubating donor spleen cells with antibodies directed at CD49d and CD62L could significantly delay the occurrence of acute GVHD. To test the potential usefulness of this treatment in BMT, we examined this therapeutic protocol in a mouse BMT model. The present mouse BMT study confirmed our previous results that incubation of donor cells with antibodies directed at CD49d and CD62L prior to infusion into the recipient can effectively delay acute GVHD, allowing the recipients to recover from the side effects of total body irradiation. This one-time treatment is easy and simple and may be modified for clinical usage.
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