ABSTRACT
A splicing mutation in VPS4B can cause dentin dysplasia type I (DD-I), a hereditary autosomal-dominant disorder characterized by rootless teeth, the etiology of which is genetically heterogeneous. In our study, dental follicle cells (DFCs) were isolated and cultured from a patient with DD-I and compared with those from an age-matched, healthy control. In a previous study, this DD-I patient was confirmed to have a loss-of-function splicing mutation in VPS4B (IVS7 + 46C > G). The results from this study showed that the isolated DFCs were vimentin-positive and CK14-negative, indicating that the isolated cells were derived from the mesenchyme. DFCs harboring the VPS4B mutation had a significantly higher proliferation rate from day 3 to day 8 than control DFCs, indicating that VPS4B is involved in cell proliferation. The cells were then replenished with osteogenic medium to investigate how the VPS4B mutation affected osteogenic differentiation. Induction of osteogenesis, detected by alizarin red and alkaline phosphatase staining in vitro, was decreased in the DFCs from the DD-I patient compared to the control DFCs. Furthermore, we also found that the VPS4B mutation in the DD-I patient downregulated the expression of osteoblast-related genes, such as ALP, BSP, OCN, RUNX2, and their encoded proteins. These outcomes confirmed that the DD-I-associated VPS4B mutation could decrease the capacity of DFCs to differentiate during the mineralization process and may also impair physiological root formation and bone remodeling. This might provide valuable insights and implications for exploring the pathological mechanisms underlying DD-I root development.
Subject(s)
Humans , ATPases Associated with Diverse Cellular Activities , Genetics , Case-Control Studies , Cell Differentiation , Genetics , Cells, Cultured , Dental Sac , Cell Biology , Dentin Dysplasia , Genetics , Pathology , Endosomal Sorting Complexes Required for Transport , Genetics , Mutation , Genetics , Osteogenesis , Genetics , RNA Splicing , GeneticsABSTRACT
<p><b>OBJECTIVE</b>To examine the effect of aqueous extracts of Scutellaria baicalensis Georgi and Radix paeoniae Alba on periodontitis mice and compare the results of the two herbs for the treatment of the periodontitis mice.</p><p><b>METHODS</b>Sixty-four SPF 12-week-old male Kunming mice were selected and randomly divided into four groups:Control group(C); Experimental periodontitis group (P):the peridontitis models in Kunming mice were prepared by wrapping silk ligature and inoculating with putative periodontopathic bacteria; Scutellaria baicalensis Georgi treatment group (SG): periodontitis was induced by the same method described above, the mice were gavaged with Scutellaria baicalensis Georgi; Radix paeoniae Alba treatment group (RG): periodontitis was induced by the same method described above, the mice were gavaged with Radix paeoniae Alba.Four mice were sacrificed at each time point of the end of 4, 6, 8 and 10 weeks in each group. The histopathological changes of periodontal tissue were observed under microscope with HE staining. The level of serum IgG1 and IgG2a was measured by enzyme-linked immunosorbent assay (ELISA) .</p><p><b>RESULTS</b>A serious inflammatory response, alveolar progressive absorption and a large number of osteoclasts were observed in the experimental periodontitis group.However, in SG and RG, the inflammation of the periodontal tissue was decreased and tissue repair was significant. The level of serum IgG2a in SG (6 week:0.934 ± 0.006, 8 week:0.743 ± 0.009, 10 week: 0.674 ± 0.008) and RG (6 week: 1.023 ± 0.032, 8 week: 0.851 ± 0.032, 10 week:0.790 ± 0.009) was significantly decreased after the mice were gavaged with the two herbs(P < 0.01). The level of serum IgG2a in SG was significantly lower than that of RG (P < 0.01). The level of serum IgG1 in SG (6 week: 0.314 ± 0.006, 8 week: 0.344 ± 0.004, 10 week: 0.367 ± 0.006) and RG (6 week: 0.287 ± 0.005, 8 week: 0.303 ± 0.058, 10 week: 0.336 ± 0.006) were significantly increased (P < 0.01). The level of serum IgG1 in SG was significantly higher than that of RG (P < 0.01).</p><p><b>CONCLUSIONS</b>Both the aqueous extracts of Scutellaria baicalensis Georgi and Radix paeoniae Alba showed therapeutic effect on periodontitis in mice.Scutellaria baicalensis Georgi was more effective than Radix paeoniae Alba.</p>
Subject(s)
Animals , Male , Mice , Aconitum , Immunoglobulin G , Metabolism , Paeonia , Periodontitis , Metabolism , Plant Extracts , Pharmacology , Scutellaria baicalensis , WaterABSTRACT
AIM: To study the possible role of nitric oxide (NO) in the development of periodontitis and the relationship between the NO concentration and the attachment loss. METHODS: Seventy- two Sprngue- Dawley rats were randomly assigned to two groups, the control group and periodontitis group. Experimental periodontitis in rats was produced by a ligature of braided silk. The nitric oxide concentration was indirectly ascertained by the concentration of nitrite (NO2-) and nitrate (NO3-)in the gingival tissue, which was assayed by spectrophotometry. The attachment loss (AL) was measured by the technology of the cellular graphics engineering research. The histopathologic change in periodontium was observed under a light microscope by using the histotomy. RESULTS: Compared to control group, the NO2-/NO3 - concentration in gingival tissue was significantly higher in periodontitis group at four weeks and eight weeks following ligation (P＜0.01). In periodontitis group, the NO2-/NO3 - concentration in gingival tissue was higher at eight weeks than that at four weeks following ligation (P＜0.01). At four weeks and eight weeks, the AL in experimental periodontitis in rats was significantly increased than that at one week after ligation ( P＜0.01); and the AL was also much higher at eight weeks than that at four weeks (P＜0.01). CONCLUSIONS: The NO2-/NO3- concentration in the gingival tissue in periodontitis group was significantly higher than that in control group. These results demonstrate that the NO2-/NO3- concentration is related to the severity of AL, and NO synthesis is very important to the process of inflammation and lesion in periodontium. Reducing NO production may be of great therapeutic value in the treatment of periodontitis.
ABSTRACT
Aim To search the effective malaria mixed gene vaccine and to explore the immune response of vaccinated host. The recombinant pcDNA3-EBA175/HRP Ⅱ was injected alone or mixed with pcDNA3-Pfs25 of Plasmodium falciparum into mice by intramuscular route. The muscle of injected parts were treated previously, e.g. injecting 50μl of 0. 5％bupivacaine into the muscle of left latter leg, with 2mm depth. To observed the changes of IgG antibody value, the splenic lymphocyte proliferation, the ratio of CD4+/CD8+ subgroups and NK cell killing activity. After injecting recombinant pcDNA3-EBA175/HRP Ⅱ alone or mixed with pcDNA3-Pfs25 into mice by intramuscular route, it showed that sera IgG value increased, the splenic T lymphocyte proliferation stimulated by specific antigen of Plasmodium falciparum increased, the ratio of CD4+/CD8+ decreased and NK cell activity increased. Enhanced immune injection could improve host's immune reaction.It is suggested intramuscular injection is an effective immune route, and mice inoculated with coding two gene recombinant alone or that mixed with sexual stage gene recombinant could all induce increased humoral and cellular immune response, and NK cell activity.