Phytocystatin CsinCPI-2 Reduces Osteoclastogenesis and Alveolar Bone Loss.

Periodontal disease (PD) is a polymicrobial chronic inflammatory condition of the supporting tissues around the teeth, leading to the destruction of surrounding connective tissue. During the progression of PD, osteoclasts play a crucial role in the resorption of alveolar bone that eventually leads to the loss of teeth if the PD is left untreated. Therefore, the development of antiresorptive therapies targeting bone-resorbing cells will significantly benefit the treatment of PD. Here, we demonstrate the inhibitory effect of CsinCPI-2, a novel cysteine peptidase inhibitor from the orange tree, on periodontitis-induced inflammation, alveolar bone loss, and osteoclast differentiation. Using the ligature-induced periodontitis model in mice, we show that treatment with CsinCPI-2 (0.8 µg/g of body weight) significantly reduced inflammatory cell infiltrate in the connective tissue and prevented the loss of alveolar bone mass (BV/TV) caused by PD, effects associated with diminished numbers of TRAP-positive multinucleated cells. Furthermore, CsinCPI-2 significantly downregulated the numbers of inflammatory cells expressing CD3, CD45, MAC387, and IL-1ß. In vitro, CsinCPI-2 inhibited RANKL-induced TRAP+ multinucleated osteoclast formation in mouse bone marrow macrophage cultures in a concentration-dependent manner. This effect was not due to cytotoxicity, as demonstrated by the MTT assay. CsinCPI-2 inhibited RANKL-induced mRNA expression of Acp5, Calcr, and Ctsk, as well as the RANKL-induced upregulation of Nfatc1, a crucial transcription factor for osteoclast differentiation. Based on our findings, CsinCPI-2 prevents bone loss induced by PD by controlling the inflammatory process and acting directly on osteoclastogenesis, suggesting an interesting potential for CsinCPI-2 in the strategy for PD treatment.

Molecular phylogenetics and phylogeography of the white-fronted capuchin (Cebus albifrons; Cebidae, Primates) by means of mtCOII gene sequences.

A total of 696 base pairs (bp) of the mitochondrial COII gene were sequenced from 118 individuals of Cebus albifrons (plus an individual of Cebus olivaceus) sampled from diverse geographical areas of Colombia, Peru, Ecuador and Brazil. These animals represented all of the C. albifrons's taxa described by Hershkovitz (1949) in Colombia and Peru (10 out of 13 subspecies are described by this author). The sequences analyzed demonstrate the existence of three well defined groups in northern Colombia (trans-Andean): malitosus, versicolor-pleei-cesarae and leucocephalus. They arose from at least, three distinct migrations from different Amazonian groups. Five different Amazonian and Eastern Llanos C. albifrons's groups (I, II, III, IV, and V) were also found. In many Amazonian localities, some of these groups live in sympatry probably by secondary expansion after their respective formations. Amazonian group I is closely related to the versicolor-pleei-cesarae group, malitosus is closely related to Amazonian group V, while leucocephalus is closely related to Amazonian group IV. Nevertheless, our genetic analysis could not resolve the genetic relationships among the main C. albifrons groups. The ρ-statistic applied to the median-joining network yielded that the major part of the temporal splits estimated occurred in the Pleistocene, reinforcing the importance of the Pleistocene refugia during the evolution of C. albifrons.