Transcriptomic profiling and the first spatial expression analysis of candidate genes in the salivary gland of the East Asian medicinal leech, Hirudo nipponia.

Hirudo nipponia, a blood-sucking leech native to East Asia, possesses a rich repertoire of active ingredients in its saliva, showcasing significant medical potential due to its anticoagulant, anti-inflammatory, and antibacterial effects against human diseases. Despite previous studies on the transcriptomic and proteomic characteristics of leech saliva, which have identified medicinal compounds, our knowledge of tissue-specific transcriptomes and their spatial expression patterns remains incomplete. In this study, we conducted an extensive transcriptomic profiling of the salivary gland tissue in H. nipponia based on de novo assemblies of tissue-specific transcriptomes from the salivary gland, teeth, and general head region. Through gene ontology (GO) analysis and hierarchical clustering, we discovered a novel set of anti-coagulant factors-i.e., Hni-Antistasin, Hni-Ghilanten, Hni-Bdellin, Hni-Hirudin-as well as a previously unrecognized immune-related gene, Hni-GLIPR1 and uncharacterized salivary gland specific transcripts. By employing in situ hybridization, we provided the first visualization of gene expression sites within the salivary gland of H. nipponia. Our findings expand on our understanding of transcripts specifically expressed in the salivary gland of blood-sucking leeches, offering valuable resources for the exploration of previously unidentified substances with medicinal applications.

Bone formation with various bone graft substitutes in critical-sized rat calvarial defect.

OBJECTIVE: This histomorphometric study compared the efficacy of a new bone graft substitute (N-HA) derived from hen eggshell, consisted of submicron scale porous hydroxyapatite structure, in the healing of 8 mm diameter critical size defects in rat calvaria. We compared N-HA alone or in combination with calcium sulfate (CS), with a commercial bone substitute, anorganic bovine bone (Bio-Oss, BO). MATERIAL AND METHODS: Critical size defects were created in calvaria of 56 adult Sprague-Dawley rats. Animals were divided into four groups and treated with (1) unfilled defects, (2) N-HA grafts, (3) BO grafts and (4) N-HA/CS grafts. The percentage of new bone formed (NB%) was evaluated histomorphometrically after 6 and 12 weeks. RESULTS: The N-HA group exhibited more new bone formation compared with other groups at 6 and 12 weeks. Histomorphometric analysis showed greater NB% in N-HA group (11.2% at 4 weeks and 19.2% at 12 weeks) compared with those in unfilled (3.9% at 6 weeks and 6.4% at 12 weeks), BO-treated (6.4% at 6 weeks and 8.2% at 12 weeks) and N-HA/CS-treated (6.3% at 6 weeks and 12.6% at 12 weeks) groups. The N-HA group showed significant differences in NB% compared with unfilled group at 6 weeks (P=0.016), unfilled and BO-treated groups at 12 weeks (P=0.001). The addition of CS did not enhance the NB% compared with defects grafted with N-HA alone. CONCLUSION: N-HA was an osteoconductive bone substitute for treating osseous defects in critical size defects of rat calvaria.

Effects of high glucose on cellular activity of periodontal ligament cells in vitro.

Periodontal ligament (PDL) cells are the most important cells in the healing of wounds and the regeneration of periodontal tissues. The response of PDL cells regarding cellular activity to high glucose concentration levels could be the key in understanding the events associated with the dental care of brittle diabetes. We studied the effect of high glucose concentration levels on the cellular activity of PDL cells from five non-diabetic patients in vitro. PDL cells were cultured for 14 days in a normal glucose medium (1100mg/l of glucose) or in a high glucose medium (4500mg/l of glucose) and a 3-(4,5-dimethylithiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay for cellular viability was also performed. In order to evaluate the differentiation of PDL cells to osteoblast-like cells, mineralized nodule formation was induced with supplemented media containing 50microg/ml of ascorbic acid, 10mM of beta-glycerophosphate and 100nM of dexamethasone for 21 days. High glucose significantly inhibited the proliferation of PDL cells and reduced the optic density of the MTT assay. Concerning the mineralized nodule formation, the percentage of the calcified area to the total culture dish of PDL cells in high glucose level was lower than that in the normal glucose medium. In conclusion, high glucose inhibits the proliferation and differentiation of PDL cells. The data provide an explanation for the delayed periodontal regeneration and healing in diabetic patients.