• Applied Microscopy
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• v.42 no.1
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• pp.27-33
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• 2012

Transient receptor potential ankyrin 1 (TRPA1), responding to noxious cold (${\leq}17^{\circ}C$) and pungent compounds, is implicated in nociception, but little is known about the coexpression of TRPA1 and other channels or receptors involved in the nociception in craniofacial regions. To address this issue, we characterized the TRPA1-immunopositive (+) neurons in the rat trigeminal ganglion (TG) and investigated their colocalization with other proteins known to be expressed in nociceptive neurons, such as transient receptor potential vanilloid (TRPV1) and $P2X_3$ receptor, using light microscopic immunofluorescence labeling method with TRPA1 and TRPV1 or $P2X_3$ antisera. The majority of TRPA1+ neurons costained for TRPV1 (TRPV1+/TRPA1+; 58.8%, 328/558) and 41.2% only expressed TRPA1 but not TRPV1. The TRPV1+/TRPA1+ neurons were small and medium sized. In addition, we investigated the colocalization of TRPA1 with $P2X_3$, a nonselective cation channel activated by ATP that may be released in the extracellular space as a result of tissue damage and inflammation. Among all TRPA1+ TG neurons, 26.1% (310/1186) costained for $P2X_3$, whereas 73.9% (876/1186) of TRPA1+ neurons did not coexpress $P2X_3$. $P2X_3$+/TRPA1+ neurons were predominantly small and medium sized. These results suggest that TRPA1+ neurons coexpressing TRPV1 or $P2X_3$ are involved in specific roles in the transmission and processing of orofacial nociceptive information by noxious cold, heat, and inflammation.

• Journal of the Korean Association of Oral and Maxillofacial Surgeons
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• v.36 no.3
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• pp.186-196
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• 2010

Introduction: The first aim of this study was to isolate the dental tissue-derived stem cells from the dental follicle (DF), dental pulp (DP), and root apical papilla (RAP) of the extracted wisdom teeth. Second was to evaluate their characterization with the expressions of transcription factors and cell surface markers. Finally, their ability of the in vitro multi-lineage differentiations into osteogenic and adipogenic cells were compared, respectively. Materials and Methods: Dental tissues, including dental follicle, dental pulp, and root apical papilla, were separated in the extracted wisdom teeth. These three dental tissues were cultured in Dulbecco’s modified Eagle’s medium (DMEM) with supplements, respectively. After passage 3, the homogeneous shaped dental tissue-derived cells were analyzed the expression of transcription factors (Oct-4, Nanog and Sox-2) and cell surface markers (CD44, CD90 and CD105) with reverse transcription polymerase chain reaction (RT-PCR) and fluorescence-activated cell sorting (FACS) analysis. In order to evaluate in vitro multi-lineage differentiations, the culture media were changed to the osteogenic and adipogenic induction mediums when the dental tissue-derived cells reached to passage 3. The characteristics of these three dental tissue-derived cells were compared with immunohistochemistry. Results: During primary culture, heterogenous and colony formatted dental tissue-derived cells were observed in the culture plates. After passage 2 or 3, homogenous spindle-like cells were observed in all culture plates. Transcription factors and mesenchymal stem cell markers were positively observed in all three types of dental tissue-derived cells. However, the quantity of expressed transcription factors was most large in RAP-derived cells. In all three types of dental tissue-derived cells, osteogenic and adipogenic differentiations were observed after treatment of specific induction media. In vitro adipogenic differentiation was similar among these three types of cells. In vitro osteogenic differentiation was most strongly and frequently observed in the RAP-derived cells, whereas rarely osteogenic differentiation was observed in the DP-derived cells. Conclusion: These findings suggest that three types of human dental tissue-derived cells from extracted wisdom teeth were multipotent mesenchymal stem cells, have the properties of multi-lineage differentiations. Especially, stem cells from root apical papilla (SCAP) have much advantage in osteogenic differentiation, whereas dental follicle cells (DFCs) have a characteristic of easy adipogenic differentiation.

• Clinical and Experimental Reproductive Medicine
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• v.37 no.3
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• pp.217-229
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• 2010

Objective: Apoptosis plays an important role for the maintenance of the normal pregnancy. Expression of X-linked inhibitor of apoptosis (XIAP) is able to effectively prevent apoptosis and controls trophoblast cells death throughout placental development, but it is still unknown in the function of XIAP in trophoblast cells exposed to hypoxic condition, which is one of the factors causing preeclampsia. Therefore, we conducted to compare XIAP expression in normal and pre-eclamptic placenta tissues and analyzed the function of XIAP in HTR-8/SVneo trophoblast cell line exposed to hypoxic condition. Methods: The expression of XIAP was analyzed in placental tissues from the following groups of patients (none underwent labor): 1) term normal placenta (n=15); 2) term with pre-eclamptic placeneta (n=15); and 3) pre-term with pre-eclamptic placenta (n=11) using semi-quantitative RT-PCR, immunohistochemistry, and Western blot. In order to evaluate the function of XIAP in HTR-8/SVneo trophoblast cells under hypoxic condition, HIF-$1{\alpha}$ plasmids, and hypoxic condtion were transfected and treated into HTR-8/SVneo trophoblast cells for 24 hours, respectively. Results: We observed that XIAP are expressed in the syncytiotrophoblasts and syncytial knot of placental villi. The expression of XIAP was significantly decreased in preeclamptic placenta tissues than in normal placenta tissues without labor (p<0.05). Furthermore, we confirmed the XIAP expression in HTR-8/SVneo trophbolast cells exposed to hypoxia was translocated from cytoplasm into nucleus and decreased XIAP by hypoxic condition induced apoptosis in HTR-8/SVneo trophoblast cells through up-regulation of pro-apoptotic proteins. Conclusion: These results suggest that the expression of XIAP is involved in placental development as well as decreased expression of XIAP by hypoxia is associated with pre-eclampsia through inducing trophoblast cells apoptosis.