Microglia are the resident immune cells of the CNS. Upon brain damage, these cells are rapidly activated and function as tissue macrophages. The first steps in this activation still remain unclear, but it is widely believed that substances released from damaged brain tissue trigger this process. In this article, we describe the effects of the blood coagulation factor thrombin on cultured rodent microglial cells. Thrombin induced a transient Ca 2ϩ increase in microglial cells, which persisted in Ca 2ϩ -free media. It was blocked by thapsigargin, indicating that thrombin caused a Ca 2ϩ release from internal stores. Preincubation with pertussis toxin did not alter the thrombin-induced [Ca 2ϩ ] i signal, whereas it was blocked by hirudin, a blocker of thrombin's proteolytic activity. Incubation with thrombin led to the production of nitric oxide and the release of the cytokines tumor necrosis factor-␣, interleukin-6, interleukin-12, the chemokine KC, and the soluble tumor necrosis factor-␣ receptor II and had a significant proliferative effect. Our findings indicate that thrombin, a molecule that enters the brain at sites of injury, rapidly triggered microglial activation.

Toll-like receptors (TLRs) are crucial pattern recognition receptors in innate immunity that are expressed in microglia, the resident macrophages of the brain. TLR2, -4, and -9 are important in the responses against Streptococcus pneumoniae, the most common agent causing bacterial meningitis beyond the neonatal period. Murine microglial cultures were stimulated with agonists for TLR1/2 (Pam 3 CSK 4 ), TLR4 (lipopolysaccharide), and TLR9 (CpG oligodeoxynucleotide) for 24 h and then exposed to either the encapsulated D39 (serotype 2) or the nonencapsulated R6 strain of S. pneumoniae. After stimulation, the levels of interleukin-6 and CCL5 (RANTES [regulated upon activation normal T-cell expressed and secreted]) were increased, confirming microglial activation. The TLR1/2, -4, and -9 agonist-stimulated microglia ingested significantly more bacteria than unstimulated cells (P < 0.05). The presence of cytochalasin D, an inhibitor of actin polymerizaton, blocked >90% of phagocytosis. Along with an increased phagocytic activity, the intracellular bacterial killing was also increased in TLR-stimulated cells compared to unstimulated cells. Together, our data suggest that microglial stimulation by these TLRs may increase the resistance of the brain against pneumococcal infections.

Purpose: Type-I collagen, the major structural protein in bone, has beneficial properties regarding bone regeneration. Little is known about the potential effects of collagen coating on orthopedic implants. method^ 3 to 6 @cm2 of lyophilized type-I collagen was absorbed on titanium rods. Six coated and uncoated pins of 0.9 mm diameter were inserted into the tibia of adult male Wistar rats for 1, 2, 4, 7, 14, and 28 days. Specimens were embedded in methacrylate-based Technovit 9100N resin. From one portion cutting and grinding sections were obtained. The implant was removed from the other half that was depolymerized, sectioned, and mounted for immunohistochemistry.Results: At day 4, the interface around the collagen-coated implants displayed a granulation tissue with higher numbers of cathepsin D-positive mononucleated cells compared to the uncoated implants @ < 0.05). Active osteoblasts, reactive for osteopontin, were increased around the collagen-coated pins at day 4 and 7 (p < 0.01). After 28 days of implantation, direct bone contact averaged 74.9% around the collagen-coated implants and 62.1% around uncoated implants (NS). The amount of newly formed bone averaged 76.3% around the collagen-coated pins and 67.8% around the uncoated pins (NS). The histomorphometric findings were confirmed by SRpCT in two specimens. Conclusions:The earlier observation of mononuclear phagocytozing cells and the earlier and higher expression of bone-specific matrix proteins suggest an increased early bone remodeling around titanium pins through collagen coating. A tendency towards increased bone formation was observed around the coated implants.

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