Effects of novel bioactive glasses on promoting remineralization of artificial dentin caries / 北京大学学报(医学版)

OBJECTIVE@#To investigate the effects of novel bioactive glasses (BG) including PSC with high phosphorus component and FBG with fluorine-doped element on promoting remineralization of artificial dentin caries.@*METHODS@#(1) BGs were used in this study as follows: PSC (10.8%P2O5-54.2%SiO2-35.0%CaO, mol.%) were synthesized using phytic acid as the phosphorus precursor through sol-gel method. FBG (6.1%P2O5-37.0%SiO2-53.9%CaO-3.0%CaF2, mol.%) and 45S5(6.0%P2O5-45.0%SiO2-24.5%CaO-24.5%Na2O, mol.%) were synthesized by traditional melt method. (2) The above BGs were soaked in simulated body fluid (SBF) for 24 hours. Then X-ray diffraction (XRD) was used to analyze the formation of hydroxyapatite (HA) crystals. (3) Prepared 1 mm thick dentin slices were soaked in 17% ethylene diamine tetraacetic acid (EDTA) for 1 week to demineralize the dentin. Then the dentin slices treated by BG were soaked in SBF for 1 week. Field emission scanning electron micro-scopy (FE-SEM) was used to observe the surface morphology of the dentin slices. (4) Four cavities were prepared to 1 mm depth in each 2 mm thick dentin slice, then were treated with lactic acid for 2 weeks to form the artificial dentin caries. Wax, mineral trioxide aggregate (MTA), PSC and FBG were used to fill four cavities as blank control group, MTA group, PSC group and FBG group respectively. Then the spe-cimens were soaked in SBF for 4 weeks. The changes of depth and density of demineralized dentin were analyzed using Micro-CT before filling and after 2 and 4 weeks filling.@*RESULTS@#(1) PSC and FBG promoted mineral formation on the surfaces of the demineralized dentin. And the speed was faster and crystallinity was higher in PSC group than the FBG and 45S5 groups. (2) The increased mineral density of artificial dentin caries in PSC group were (185.98 ± 55.66) mg/cm3 and (213.64 ± 36.01) mg/cm3 2 and 4 weeks after filling respectively, which were significantly higher than the control group [(20.38 ± 7.55) mg/cm3, P=0.006; (36.46 ± 10.79) mg/cm3, P=0.001]. At meanwhile, PSC group was also higher than MTA group [(57.29 ± 10.09) mg/cm3; (111.02 ± 22.06) mg/cm3], and it had statistical difference (P=0.015; P=0.006). The depth of remineralized dentin in PSC group were (40.0 ± 16.9) μm and (54.5 ± 17.8) μm 2 and 4 weeks respectively, which were also statistically different from the control group (P =0.010;P=0.001). There were no statistical differences between the control group and MTA group. The above effects of FBG group were between PSC and MTA.@*CONCLUSION@#PSC has advantages in the speed, quality and depth of mineral deposition in the demineralized layer of artificial dentin caries. It would be expected to be an ideal material to promote the remineralization of dentin caries.

Avaliação microtomográfica e histomorfométrica do processo de reparo de defeitos ósseos em calvária de coelhos tratados com diferentes materiais de enxerto / Microtomographic and histomorphometric evaluation of bone repair in rabbit cranial defects treated with different graft materials

Um dos grandes desafios para o tratamento de defeitos ósseos extensos na região bucomaxilofacial têm sido o desenvolvimento de um biomaterial substituto ósseo ao enxerto autógeno. No presente trabalho avaliou-se a formação óssea e a biodegrabilidade do osso desproteinizado bovino Bio-Oss® e do seu similar GenOx Inorg® e da cerâmica bifásica GenPhos® XP no processo de reparo de defeitos ósseos cranianos em coelhos, comparativamente ao osso autógeno (controle positivo) e coágulo sanguíneo (controle negativo). Foram realizados cirurgicamente defeitos bilaterais de 8-mm de diâmetro nos ossos parietais de 39 coelhos. A seguir os defeitos foram preenchidos aleatoriamente com 0,1cm3 de material ou coágulo conforme cada grupo de tratamento. Após os períodos de 4, 8 e 24 semanas os crânios foram coletados, analisados no microtomógrafo e processados histologicamente. O percentual de volume do defeito ocupado pelo material e osso neoformado foi avaliado pela microtomografia e histomorfometria, enquanto que, para a medula óssea, tegumento e tecido conjuntivo, apenas pela análise histomorfométrica. Os resultados quantitativos obtidos foram comparados estatisticamente pela ANOVA a dois critérios (período e tratamento) e teste de Tukey com p<0,05. A intensidade da associação linear dos dados microtomográficos e histomorfométricos avaliada pelo coeficiente de correlação de Pearson, mostraram correlação moderada a forte. Nos períodos iniciais de reparo (30 e 60 dias), os defeitos tratados com Bio-Oss®, GenOx® Inorg e GenPhos® XP apresentaram manutenção do volume do material enxertado (Vvi médio de 34% ) e formação óssea menor e mais imatura em relação grupo autógeno (Vvi = 22% vs. 32% no grupo autógeno). No período mais tardio (180 dias) a quantidade de formação óssea foi estatisticamente similar nos grupos Bio-Oss® (Vvi = 27%), GenOx® Inorg (Vvi = 26%) e GenPhos® XP (Vvi = 20%) porém, o GenOx® Inorg promoveu a formação de um tecido ósseo mais organizado e com maior acúmulo de biomaterial+osso+medula óssea (Vvi = 67,9%) comparado ao GenPhos® XP (Vvi =58,9%) e Bio Oss (Vvi = 55,6%) mas, inferior ao do enxerto autógeno (Vvi = 78%). Os resultados aqui obtidos permitem concluir que o osso autógeno promove rápida formação e maturação óssea, porém não consegue promover o reestabelecimento completo da díploe removida cirurgicamente. Os materiais BioOss, GenOx® Inorg e GenPhos® XP são excelentes materiais osteocondutores levando a formação óssea em toda extensão do defeito, sendo o GenOx® Inorg o que apresenta menor grau de reabsorção e maior e melhor preenchimento do defeito.(AU)

One major challenge for treatment of critical size defects in maxillofacial region has been the development of a substitute biomaterial to the autogenous bone grafts. In present study we evaluated the bone formation and biodegradability of deproteinized bovine bone Bio-Oss® and GenOx® Inorg, and biphasic calcium phosphate GenPhos XP® during bone repair process in rabbits cranial defects compared to autogenous bone (positive control) and blood clot (negative control). In parietal bone of 39 rabbits were made bilateral 8-mm diameter defects, which were filled randomly with 0,1cm3 material or clot as each treatment group. After periods of 4, 8 and 24 weeks skulls of animals were collected, analyzed the MicroCT scanner and histologically processed. The percentage of defect volume occupied by biomaterial and new-formed bone were assessed by histomorphometry and microtomography, while the bone marrow, connective tissue and tegument only by first analysis. The quantitative data were compared by two-way ANOVA analysis (time and treatment) and Tukey's test at p <0.05. The intensity of the linear association of MicroCT and morphometric data evaluated by the Pearson correlation coefficient, showed moderate to strong correlation. In the early repair periods (30 and 60 days), the defects treated with Bio- Oss, GenOx® Inorg and GenPhos® XP showed maintenance of the graft material volume (average Vvi of 34%) and lower and more immature bone compared autograft group (Vvi = 22% vs. 32% in the autograft group). In the later period (180 days) the amount of bone formation was statistically similar to the groups Bio-Oss® (Vvi = 27 %), GenOx® Inorg (Vvi = 26%) and GenPhos® XP (Vvi = 20%) however, the bone formation in GenOx® Inorg was more organized and with greater accumulation of particles + bone tissue + bone marrow (Vvi = 67.9%), when compared to GenPhos® XP (Vvi = 58.9%) and Bio-Oss® (Vvi = 55.6%) but lower than the autograft (Vvi = 78%). It was concluded that the autogenous bone promotes rapid bone formation and maturation, but cannot promote the complete reestablishment of diploe surgically removed. The Bio-Oss®, GenOx® Inorg and GenPhos® XP are excellent osteoconductive materials leading to bone formation in the full extent of the defects, and the GenOx® Inorg showing less absorption promotes more and better defect filling.(AU)

Repair of bone cavities in dog's mandible filled with inorganic bovine bone and bioactive glass associated with platelet rich plasma

The aim of this study was to evaluate the effect of platelet rich plasma (PRP) associated to bovine inorganic bone (Bio-Oss®; Geistlich) or bioactive glass (Bio-Gran®; Orthovita, Implant Innovations) on bone healing. Bone cavities were prepared in both sides of the mandible of 4 adult male dogs. The cavities were divided into 4 groups according to the filling material as follows: control, PRP, PRP/Bio-Oss, PRP/Bio-Gran. The animals were sacrificed after 120 days and histological and histomorphometrical analysis was performed. The control group showed 80.6 percent of bone formation in the longitudinal sections at 6 mm depth and 83.7 percent at 13 mm depth. The transverse sections displayed 74.2 percent at both 6 and 13 mm depths. The PRP group showed 21.1 percent of bone formation in the longitudinal sections at 6 mm depth, and 23.1 percent at 13 mm depth. The transverse sections presented 28.98 percent of bone formation at 6 mm depth and 41.2 percent at 13 mm depth. The PRP/Bio-Gran group showed 25.1 percent of bone formation in the longitudinal sections at 6 mm depth and 30.4 percent at 13 mm depth. In the transverse sections, the bone formation was 43.0 percent at 6 mm depth and 39.7 percent at 13 mm depth. The PRP/Bio-Oss group showed 35.5 percent of bone formation in the longitudinal sections at 6 mm depth and 42 percent at 13 mm depth. In the transversal sections, the bone formation was 26.8 percent and 31.2 percent at the depths of 6 and 13 mm, respectively. PRP alone or associated with bovine inorganic bone or bioglass had no significant effect in bone healing.

O objetivo deste trabalho foi avaliar os efeitos do PRP associado ao osso bovino inorgânico ou vidro bioativo no processo de reparo ósseo. Foram utilizados 4 cães adultos, onde foram preparadas cavidades ósseas em região mandibular dos dois lados. As cavidades foram divididas em 4 grupos de acordo com o material utilizado: controle, PRP, PRP/Bio-Oss®, PRP/Bio-Gran®. O sacrifício dos animais foi realizado 120 dias depois do procedimento cirúrgico e análises histológicas e histomorfométricas foram realizadas. O grupo controle demonstrou 80,6 por cento de formação óssea no corte longitudinal com profundidade de 6 mm e 83,7 por cento no corte longitudinal com profundidade de 13 mm. Nos cortes transversais de 6 e 13 mm de profundidade mostrou 74,2 por cento de formação óssea. O grupo PRP demonstrou 21,1 por cento de formação óssea no corte longitudinal com profundidade de 6 mm e 23,1 por cento no corte longitudinal com profundidade de 13 mm. Nos cortes transversais de 6 mm de profundidade mostrou 28,98 por cento e 41,2 por cento nos de 13 mm. O grupo PRP/bio-Gran® demonstrou 25,1 por cento de formação óssea no corte longitudinal com profundidade de 6 mm e 30,4 por cento no corte longitudinal com profundidade de 13 mm. Nos cortes transversais de 6 mm de profundidade mostrou 43,0 por cento de formação óssea e 39,7 por cento nos de 13 mm. Para o grupo PRP/Bio-Oss® nos cortes longitudinais obtivemos 35,5 por cento para os cortes de 6 mm e 42 por cento para os 13 mm. Nos cortes transversais a formação óssea encontrada foi de 26,8 por cento e 31,2 por cento paras as profundidades de 6 e 13 mm respectivamente. Conclui-se que o uso isolado associado do PRP com o osso bovino inorgânico ou vidro bioativo não possui um efeito significativo no processo de reparo ósseo.

Subcutaneous tissue reaction to castor oil bean and calcium hydroxide in rats

Castor oil bean cement (COB) is a new material that has been used as an endodontic sealer, and is a candidate material for direct pulp capping. OBJECTIVE: The purpose of this study was to evaluate the biocompatibility of a new formulation of COB compared to calcium hydroxide cement (CH) and a control group without any material, in the subcutaneous tissue of rats. MATERIAL AND METHODS: The materials were prepared, packed into polyethylene tubes, and implanted in the rat dorsal subcutaneous tissue. Animals were sacrificed at the 7th and 50th days after implantation. A quantitative analysis of inflammatory cells was performed and data were subjected to ANOVA and Tukey's tests at 5 percent significance level. RESULTS: Comparing the mean number of inflammatory cells between the two experimental groups (COB and CH) and the control group, statistically significant difference (p=0.0001) was observed at 7 and 50 days. There were no significant differences (p=0.111) between tissue reaction to CH (382 inflammatory cells) and COB (330 inflammatory cells) after 7 days. After 50 days, significantly more inflammatory cells (p=0.02) were observed in the CH group (404 inflammatory cells) than in the COB group (177 inflammatory cells). CONCLUSIONS: These results demonstrate that the COB cement induces less inflammatory response within long periods.

[Effect of vitamin-mineral supplementation on oxidative stress and plasma cytokine response after strenuous training in female elite swimmers]

This study aims at investigating the effect of vitamin-mineral supplementation on oxidative stress and plasma cytokine response after strenuous training periods in female elite swimmers. Twenty-four elite female swimmers volunteered to participate in this study and were randomly divided into two groups, the experimental [Vitamin-mineral supplemented] and the control [Placebo]. Both groups were in a monthly swimming programs, 3 times a week, for a total of 4 weeks and swimming, almost 3.5 to 4 km/d. Blood sampling was done before and after the training period to assess inflammatory cytokines such as IL-6 and TNF-alpha, and also MDA. 100 m crawl records were measured at the beginning and the end of the training period. Results showed that inflammatory cytokines decreased significantly in the vitamin-mineral supplemented group, and MDA decreased, though not significantly, in this group. There was no significant change between the groups. No significant change was observed in swimming performance in either groups. In conclusion, ROS was found to affect exercise-induced cytokine production, in which vitamin-mineral supplementation was found to play an effective role

Biodisponibilidade de minerais em ratos alimentados com frutanos do tipo inulina / Mineral bioavailability in rats fed inulin-type fructans

Neste estudo foi avaliado o efeito de frutanos do tipo inulina (FTI) na biodisponibilidade de minerais em ratos, com particular enfoque para o Ca e o Fe. Foram realizados dois ensaios: no primeiro utilizou-se um modelo fatorial 2×2×2, com dois níveis de Fe (0 e 35 mg/kg), Ca (0 e 5 g/kg) e FTI (0 e 100 g/kg) nas rações; no segundo, utilizou-se um modelo de repleção de Hb em ratos anêmicos. As fontes de FTI foram a Raftilose P95 (ensaios 1 e 2), fonte purificada, e a farinha de yacón (FY; ensaio 2); os sais de Fe utilizados foram o Fe4(P2O7)3 (ensaios 1 e 2) e o FeSO4 microencapsulado com alginato (ensaio 2). No primeiro ensaio foram avaliados parâmetros bioquímicos de Fe, concentração de minerais nos ossos, fígado e baço, e parâmetros de resistência e histomorfometria óssea. Após 33 d, não foram observados efeitos na retenção óssea de Ca, porém observou-se maior volume trabecular ósseo e parâmetros histomorfométricos de formação óssea nos animais deficientes em Fe alimentados com os FTI. No segundo ensaio foram determinados parâmetros hematológicos, de biodisponibilidade de Fe, e parâmetros intestinais (balanço mineral; pH, ácidos graxos de cadeia curta e espécies de Fe no conteúdo intestinal). A FY recuperou os animais da deficiência de Fe mais rapidamente do que a Raftilose. Esse efeito ocorreu em função de alterações intestinais provocadas pelo aumento da fermentação desses carboidratos, que afetou a solubilidade do Fe da dieta. Concluiu-se que os FTI aumentaram a biodisponibilidade de minerais, possivelmente por influência da concentração do mineral e pelas interações entre os minerais presentes na ração, pelo tempo de ensaio, pela matriz alimentar, e pelo modelo experimental utilizado

This study evaluated the effect of inulin-type fructans (ITF) on mineral bioavailability in rats, focusing mainly on Ca and Fe. Two experiments were carried out: the first one considered a 2×2×2 factorial model using two levels of Fe (0 and 35 mg/kg), Ca (0 and 5 g/kg) and ITF (0 and 100 g/kg) in the diets; the second experiment considered a Hb repletion model in anemic rats. Raftilose P95 (experiments 1 and 2; purified source) and yacon flour (YF; experiment 2) were used as ITF sources; the Fe salts used were Fe4(P2O7)3 (experiments 1 and 2) and microencapsulated FeSO4 with alginate (experiment 2). In the first experiment, Fe biochemical parameters, mineral concentration in the bones, liver and spleen, bone strength and histomorphometrical parameters were evaluated. After 33 d, no effects on bone retention were observed, but a larger bone volume and larger bone-formation parameters were observed on Fe-deficient animals fed with ITF. In the second experiment, hematological, Fe bioavailability and intestinal parameters (mineral balance, pH, short-chain fatty acids and Fe species in the intestinal content) were determined. Fe-deficient animals fed with YF recovered more rapidly than those fed with Raftilose. This effect took place as a function of intestinal alterations caused by the increased fermentation of these carbohydrates, which affected dietary Fe solubility. It can be concluded that ITF increased the bioavailability of minerals, possibly due to the influence of mineral concentration and interactions between minerals in the diet, experiment duration, food matrix and experimental model used

Combined effects of enhanced ultraviolet-B radiation and mineral nutrients on growth, biomass accumulation and yield characteristics of two cultivars of Vigna radiata L.

In a field experiment, the effect of enhanced UV-B radiation (simulating 20% ozone depletion at Allahabad, 20 degrees 47' N latitude) was studied on two cultivars of Vigna radiata L. with various levels of mineral nutrients (N and P). Study showed decrease in total biomass accumulation, harvest index, RSR and yield after exposure with enhanced level of UV-B. RGR and CGR also showed decline after exposure with UV-B. Application of recommended dose of mineral nutrients alleviated the deleterious effect of UV-B and increased plant dry matter vis a vis yield. Both cultivars showed sensitivity to UV-B but cultivar Malviya Janpriya was more responsive to UV-B than Malviya Jyoti.

Immunostimulatory effects of anionic alkali mineral complex solution Barodon in porcine lymphocytes

The anionic alkali mineral complex solution, Barodon (Barodon-S.F. Corp., Korea), was evaluated for its effectiveness as a nonspecific immunostimulator in pigs. The effects of Barodon were determined by analysis of feed efficiency, growth rate, and phenotype of leukocyte subpopulations using monoclonal antibodies specific to porcine leukocyte differentiation antigens and flow cytometry (FC). The study was focused to investigate the change in proportion of the CD4+CD8+ double positive T lymphocyte subpopulation (dpp) which exists uniquely in pigs. In addition, the mitogen-stimulated lymphoproliferative response, tissue distribution in lymphoid organs and the adjuvant effect of Barodon on hog cholera vaccine efficiency were determined. The study has revealed the average daily gain rates and feed conversion rates were significantly (p<0.05) improved in either group of pigs fed with 0.05% Barodon-spray feed (Tx-1) or pigs fed with 3% Barodon-fermented feed (Tx-2) in comparison with group of pigs fed with feed containing no Barodon (control). The proportion of cells expressing CD4+ antigen in Barodon-treated group increased from 3 weeks posttreatment and was significantly higher (p<0.05) than that of control at 8 weeks posttreatment. Particularly, the significantly higher proportion was maintained from 8 weeks through 13 weeks posttreatment in Tx-1 group (p<0.05). The proportion of cells expressing CD8+ antigen was significantly higher at 3 weeks posttreatment in Tx-2 (p<0.01). Proportion of MHC class II-expressing cells was significantly higher in Tx-1 and Tx-2 group at 11 weeks and 8 weeks posttreatment (p<0.05), respectively. In addition, the proportion of Non T/Non B (N) cells was also significantly higher in Tx-2 at 3 weeks posttreatment (p<0.01) and maintained to 13 weeks posttreatment (p<0.1). Between Barodon-treated groups, the proportion of MHC class II-expressing cells was observed to be larger in Tx-2 than Tx-1 from 3 weeks to 8 weeks posttreatment (p<0.05). However, there were no significant difference in the proportions of CD2+ cells, B cells, monocytes and granulocytes between Barodon-treated and control group during the experiment. Dual-color FC analysis, study has revealed an increased proportion of dpp present in lymphocytes obtained from peripheral blood (PB) and mesenteric lymph node (MLN) of Barodon-treated group at 8 and 11 weeks posttreatment. The proportion of dpp in PB was 27.5% and 32.1% in Tx-1 and Tx-2, respectively, but only 2.2% in control group at 8 weeks posttreatment. In MLN, the proportion was 45.1% and 52.1% in Tx-1 and Tx-2, respectively, otherwise 16.5% in control group at 8 weeks posttreatment. The mitogen-stimulated activity was significantly higher in Tx-1 than in the control group at 11 weeks posttreatment when cells were stimulated with Con A and PHA, respectively (p<0.01). Also, Con A-, PHA and PWM-stimulated activity was significantly higher in Tx-2 than in the control group at the same time (p<0.05). The tissue distribution of CD4+, CD8+ and CD4+CD8+ dpp in MLN and spleen was significantly larger in Tx-1 and Tx-2 than in the control group (p<0.01). Also, a larger proportion of dpp was observed in Tx-2 than Tx-1 in spleen between Barodon-treated groups (p<0.01). In conclusion, the study has demonstrated that Barodon had an immunostimulatory effect on pigs through proliferation and activation of porcine immune cells, specially CD4+CD8+ dpp lymphocytes.

Propagation of Mycobacterium lepraemurium on supplemented minimal medium and its experimental pathogenesis.

The splenic tissue of a mouse experimentally infected with M. lepraemurium (Hawaiian strain, M-65) and developing 'rat leprosy', yielded a pure culture of an acid - fast bacterium having all the characteristics of M. lepraemurium on mineral salt minimal medium supplemented with simple sources of C and N, e.g., NH4 -salts, liquid paraffin, urea, gelatin etc. This could be maintained, by serial passages in vitro with good growth. Its indefinite propagation with tissue - free washed, small inoculum on complex media including Ogawa medium was difficult, and its serial sub-culture was practically impossible. The in vitro isolate from supplemented minimal medium could produce pathological lesions in mice typical of rat leprosy.