Fig 1.
A: computerized reeling machine for spider silk harvest [30]. The spider is carefully fixed underneath a compress with needles placed between the spiders’ legs and in front of the head, indicated by a star. B shows a female Nephila edulis spider in the department’s animal facility. In c, the apparatus for fibrin manufacturing can be seen. D shows the fibrin sheath containing spider silk (reeled between the metal frame). E is a scheme of the implant. It shows the dense fibrin tube containing the longitudinally orientated spider silk which is held in position by a fibrin filling.
Table 1.
Modified score for the evaluation of animals wellbeing.
Fig 2.
Spider silk (a) and fibrin scaffold (b) were seeded with olfactory ensheathing cells. Vital cells are green, while apoptotic cells (barely visible) appear red. Orange-red spider silk autofluorescence is faintly visible in a. Scale bars pertain to 500 μm.
Fig 3.
In-vitro examination of spinal cord tissue on spider silk.
A-f Scanning electron microscopy of spinal cord tissue explant on spider silk after 21 days of cultivation. The tissue explant can be seen on the right site of a. Cells using the spider silk as guidance are documented in several magnifications. G-k Immuno-fluorescence staining of spinal cord tissue explant on spider silk after 21 days of cultivation. G Neuron-specific class III beta-tubulin (Tuj-1) indicating neuronal cell bodies and axons (green). H Nestin, intermediate filament protein, as a marker for neural stem cells (red). I CX3C chemokine receptor 1 (CX3CR1) indicating microglial-cells (green). J Myelin basic protein (MBP) indicating oligodendrocytes (green). K Glial fibrillary acidic protein (GFAP), intermediate filament protein indicating astrocytes and ependymal cells (green). Nuclei were counterstained with DAPI and are blue in all pictures.
Fig 4.
Foreign body reaction in-vivo.
A (H&E) and b (EvG) display overview images of the spinal cord, which can be partitioned into three morphological sections. The foreign-body granuloma (b—arrow) and the migration of fibroblasts (b–arrowhead) are visible in b. In c (H&E) polynuclear foreign-body giant cells (c–arrow), epithelioid cells (c- plus), and the fibrin part of the initially placed implant (c–star) can be seen inside the foreign-body granuloma (b–arrow). D (EvG) is a magnification of b showing collagen (d–star) formed by migrating fibroblasts as a part of the gliomesenchymal scaring, starting from the arachnoid (d—plus). Hemosiderophages (e—arrow), indicating the old bleeding caused by the surgical procedure, and foamy macrophages (e–star) are visible in e (Prussion blue). Microvascular proliferation (f–arrows), as a part of the spinal cord infarction area, can be seen in f (H&E). G shows a magnification of the area marked in b with a star, stained for the glial fibrillary acidic protein, specifically expressed in astrocytes. No astrocytes are located beyond the transition to the implant (g–star). H and i were stained for tubulin, indicating neuronal bodies and axons. In h degenerated axons (h- arrow), in the form of Wallerian degeneration are visible, while i shows healthy axons, as a comparison, of the control.