Rabbit Anti-Human PLG (heavy chain A short form, Cleaved-Val98) polyclonal antibody

Ethnopharmacological relevance: The bulbs and flowers of plants from the Lilium genus have historically been used in Asian and Greco-Roman medicine to treat burns and promote skin healing. Aim of the study: To evaluate a steroidal glycoalkaloid isolated from Easter lily bulbs for its potential wound healing promoting properties. Materials and methods: A lily-derived steroidal glycoalkaloid (LSGA), (22R, 25R)-spirosol-5-en-3 beta-yl O-alpha-L-rhamnopyranosyl-(1 -> 2)-beta-D-glucopyranosyl-(1 -> 4)-beta-D-glucopyranoside, was isolated from Easter lily bulbs, and its structure was confirmed by LC-MS and NMR spectrometry. LSGA effects on wound scratch closure were evaluated in a primary human dermal fibroblast cell culture, and the changes in gene expression profiles were quantitated using an 84 wound-related gene qPCR microarray. Results: LSGA promoted migration of dermal fibroblasts into the wounded area. The treatment was associated with a rapid upregulation of early inflammatory (CD40LG, CXCL11, IFNG, IL10, IL2 and IL4), cell growth (CSF3 and TNF) and remodeling (CTSG, F13A1, FGA, MMP and PLG) genes both in the wounded and unwounded cells treated with LSGA. A selective decrease in gene expression profiles associated with inflammatory (CXCL2 and CCL7) and remodeling (MMP7 and PLAT) phases was observed in wounded cells treated with LSGA, in contrast to the wounded cells (control). Conclusion: This study demonstrates that a glycoalkaloid present in lilies promoted fibroblast migration in vitro and affected inflammatory, remodeling and growth factor gene expression. The decreases in expression of key genes may impact the wound healing process, possibly contributing to an earlier end of the inflammatory response and shortening the early phases of model tissue reconstitution. The results of this preliminary investigation may provide a basis for the historical use of lily bulbs to promote dermal healing after injury.

Surface modification with affinity ligands capable of capturing bioactive molecules in situ is a widely used strategy for developing biofunctional materials. However, many bioactive molecules, for example zymogens, exist naturally in a "quiescent'' state, and become active only when "triggered'' by specific activators. In the present study, in situ activation of a surface-integrated zymogen was achieved by introducing affinity ligands for both the zymogen and its activator. Specifically a dual affinity surface was designed for the integration of plasminogen (Plg) and tissue plasminogen activator (t-PA). This surface was expected to have plasmin-generating and, therefore, fibrinolytic properties. A polyurethane surface was modified with a copolymer of 2-hydroxyethyl methacrylate and 1-adamantan-1-ylmethyl methacrylate poly(HEMA-co-AdaMA). The affinity ligands, ARMAPE peptide (for t-PA) and e-lysine-containing beta-cyclodextrin (beta-CD-(Lys)(7)) (for Plg), were attached in sequence via covalent bonding and host-guest interactions, respectively. The resulting surfaces were shown to have high binding capacities for both t-PA and Plg while resisting nonspecific protein adsorption. Pre-loading with t-PA followed by Plg uptake from plasma generated plasmin and thus endowed the surface with fibrinolytic activity. In general the incorporation of dual affinity ligands to achieve surface-promoted bioactivity is a promising approach for the development of biofunctional materials. The method reported herein for the sequential attachment of plasminogen and t-PA affinity ligands can be extended to systems of multiple ligands generally.