Plastin 3 influences bone homeostasis through regulation of osteoclast activity.

Over 200 million people suffer from osteoporosis worldwide, one third of which will develop osteoporotic bone fractures. Unfortunately, no effective cure exists. Mutations in plastin 3 (PLS3), an F-actin binding and bundling protein, cause X-linked primary osteoporosis in men and predisposition to osteoporosis in postmenopausal women. Moreover, the strongest association so far for osteoporosis in elderly women after menopause was connected to a rare SNP in PLS3, indicating a possible role of PLS3 in complex osteoporosis as well. Interestingly, 5% of the general population are overexpressing PLS3, with yet unknown consequences. Here, we studied ubiquitous Pls3 knockout and PLS3 overexpression in mice and demonstrate that both conditions influence bone remodeling and structure: while Pls3 knockout mice exhibit osteoporosis, PLS3 overexpressing mice show thickening of cortical bone and increased bone strength. We show that unbalanced PLS3 levels affect osteoclast development and function, by misregulating the NFκB pathway. We found upregulation of RELA (NFκB subunit p65) in PLS3 overexpressing mice-known to stimulate osteoclastogenesis-but strikingly reduced osteoclast resorption. We identify NFκB repressing factor (NKRF) as a novel PLS3 interactor, which increasingly translocates to the nucleus when PLS3 is overexpressed. We show that NKRF binds to the NFκB downstream target and master regulator of osteoclastogenesis nuclear factor of activated T cells 1 (Nfatc1), thereby reducing its transcription and suppressing osteoclast function. We found the opposite in Pls3 knockout osteoclasts, where decreased nuclear NKRF augmented Nfatc1 transcription, causing osteoporosis. Regulation of osteoclastogenesis and bone remodeling via the PLS3-NKRF-NFκB-NFATC1 axis unveils a novel possibility to counteract osteoporosis.

Biomechanics and biocompatibility of woven spider silk meshes during remodeling in a rodent fascia replacement model.

OBJECTIVE: The aim of this study was to investigate biomechanical and immunogenic properties of spider silk meshes implanted as fascia replacement in a rat in vivo model. BACKGROUND: Meshes for hernia repair require optimal characteristics with regard to strength, elasticity, and cytocompatibility. Spider silk as a biomaterial with outstanding mechanical properties is potentially suitable for this application. METHODS: Commercially available meshes used for hernia repair (Surgisis and Ultrapro) were compared with handwoven meshes manufactured from native dragline silk of Nephila spp. All meshes were tied onto the paravertebral fascia, whereas sham-operated rats were sutured without mesh implantation. After 4 or 14 days, 4 weeks, and 4 or 8 months, tissue samples were analyzed concerning inflammation and biointegration both by histological and biochemical methods and by biomechanical stability tests. RESULTS: Histological sections revealed rapid cell migration into the spider silk meshes with increased numbers of giant cells compared with controls with initial decomposition of silk fibers after 4 weeks. Four months postoperatively, spider silk was completely degraded with the formation of a stable scar verified by constant tensile strength values. Surgisis elicited excessive stability loss from day 4 to day 14 (P < 0.001), with distinct inflammatory reaction demonstrated by lymphocyte and neutrophil invasion. Ultrapro also showed decreasing strength and poor elongation behavior, whereas spider silk samples had the highest relative elongation (P < 0.05). CONCLUSIONS: Hand-manufactured spider silk meshes with good biocompatibility and beneficial mechanical properties seem superior to standard biological and synthetic meshes, implying an innovative alternative to currently used meshes for hernia repair.

Artificial skin--culturing of different skin cell lines for generating an artificial skin substitute on cross-weaved spider silk fibres.

BACKGROUND: In the field of Plastic Reconstructive Surgery the development of new innovative matrices for skin repair is in urgent need. The ideal biomaterial should promote attachment, proliferation and growth of cells. Additionally, it should degrade in an appropriate time period without releasing harmful substances, but not exert a pathological immune response. Spider dragline silk from Nephila spp meets these demands to a large extent. METHODOLOGY/PRINCIPAL FINDINGS: Native spider dragline silk, harvested directly out of Nephila spp spiders, was woven on steel frames. Constructs were sterilized and seeded with fibroblasts. After two weeks of cultivating single fibroblasts, keratinocytes were added to generate a bilayered skin model, consisting of dermis and epidermis equivalents. For the next three weeks, constructs in co-culture were lifted on an originally designed setup for air/liquid interface cultivation. After the culturing period, constructs were embedded in paraffin with an especially developed program for spidersilk to avoid supercontraction. Paraffin cross-sections were stained in Haematoxylin & Eosin (H&E) for microscopic analyses. CONCLUSION/SIGNIFICANCE: Native spider dragline silk woven on steel frames provides a suitable matrix for 3 dimensional skin cell culturing. Both fibroblasts and keratinocytes cell lines adhere to the spider silk fibres and proliferate. Guided by the spider silk fibres, they sprout into the meshes and reach confluence in at most one week. A well-balanced, bilayered cocultivation in two continuously separated strata can be achieved by serum reduction, changing the medium conditions and the cultivation period at the air/liquid interphase. Therefore spider silk appears to be a promising biomaterial for the enhancement of skin regeneration.