Identification of PCPE-2 as the endogenous specific inhibitor of human BMP-1/tolloid-like proteinases.

BMP-1/tolloid-like proteinases (BTPs) are major players in tissue morphogenesis, growth and repair. They act by promoting the deposition of structural extracellular matrix proteins and by controlling the activity of matricellular proteins and TGF-ß superfamily growth factors. They have also been implicated in several pathological conditions such as fibrosis, cancer, metabolic disorders and bone diseases. Despite this broad range of pathophysiological functions, the putative existence of a specific endogenous inhibitor capable of controlling their activities could never be confirmed. Here, we show that procollagen C-proteinase enhancer-2 (PCPE-2), a protein previously reported to bind fibrillar collagens and to promote their BTP-dependent maturation, is primarily a potent and specific inhibitor of BTPs which can counteract their proteolytic activities through direct binding. PCPE-2 therefore differs from the cognate PCPE-1 protein and extends the possibilities to fine-tune BTP activities, both in physiological conditions and in therapeutic settings.

Combination of Traction Assays and Multiphoton Imaging to Quantify Skin Biomechanics.

An important issue in tissue biomechanics is to decipher the relationship between the mechanical behavior at macroscopic scale and the organization of the collagen fiber network at microscopic scale. Here, we present a protocol to combine traction assays with multiphoton microscopy in ex vivo murine skin. This multiscale approach provides simultaneously the stress/stretch response of a skin biopsy and the collagen reorganization in the dermis by use of second harmonic generation (SHG) signals and appropriate image processing.

Development of human corneal epithelium on organized fibrillated transparent collagen matrices synthesized at high concentration.

Several diseases can lead to opacification of cornea requiring transplantation of donor tissue to restore vision. In this context, transparent collagen I fibrillated matrices have been synthesized at 15, 30, 60 and 90 mg/mL. The matrices were evaluated for fibril organizations, transparency, mechanical properties and ability to support corneal epithelial cell culture. The best results were obtained with 90 mg/mL scaffolds. At this concentration, the fibril organization presented some similarities to that found in corneal stroma. Matrices had a mean Young's modulus of 570 kPa and acellular scaffolds had a transparency of 87% in the 380-780 nm wavelength range. Human corneal epithelial cells successfully colonized the surface of the scaffolds and generated an epithelium with characteristics of corneal epithelial cells (i.e. expression of cytokeratin 3 and presence of desmosomes) and maintenance of stemness during culture (i.e. expression of ΔNp63α and formation of holoclones in colony formation assay). Presence of cultured epithelium on the matrices was associated with increased transparency (89%).

Study of dural suture watertightness: an in vitro comparison of different sealants.

BACKGROUND: CSF leakages constitute a major complication of intradural procedures, especially for posterior fossa and skull base surgery. Dural suture watertightness is a decisive issue, and neurosurgeons routinely use different products to reinforce their dural closure. We have designed an experimental system capable of testing CSF leak pressure levels in order to compare two types of sutures in vitro and particularly four different sealants. METHODS: Twenty-five fresh human cadaveric dural samples were removed and prepared for testing in a pressure chamber system connected to a hydraulic pressure motor. CSF leak levels were objectively registered. First, simple interrupted stitches were compared to running simple closure on 50-mm linear suture. Secondly, four sealants (two sealants/glues, Bioglue®, Duraseal®; two haemostatics, Tachosil®, Tissucol®) were tested. Statistical analysis was performed with paired Student's t-test. RESULTS: No significant difference between interrupted closure and running suture was observed (p = 0.079). All sealants increased the watertightness of the suture significatively. However, comparison of the means of the differences for each product revealed large variations. In the conditions of our experiment, one sealant (Duraseal®) and one haemostatic (Tachosil®) seemed to show better results. We observed two different types of leakage: at the dura-sealant interface and through the sealant itself. CONCLUSIONS: We have developed an experimental device capable of testing dural closure watertightness. Interrupted stitch suturing seemed no different from running simple closure. On the contrary, the sealants tested show different watertightness capacities.