Topophysis and Growth Regulators Effects on Buds and Zygotic Embryos Regeneration of African Bush Mango Tree Cultured In vitro

Background: African bush mango tree is an important fruit plant with high nutritional, medicinal, and commercial values. However, its seedling system remained a deep understanding. This study aimed to evaluate the effect of topophysis and growth regulators on the reactivity of different types of buds and zygotic embryos of wild mango. Methodology: Ripe fruits from two local varieties (Wossro and Sissro) were pulped. The nuts were extracted and dried for one week under greenhouse. Zygotic embryos were excised from nuts and disinfected with the bleach solution (NaClO 10% with 8% active chloride) for 10 min following by three successive rinsing using sterilized water. The second set of nuts was cultivated for under greenhouse in the polybags containing the sand. One month later, buds from different positions (apical, axillary and cotyledonary) were excised and disinfected with NaClO 10% for 10 min follow by the immersion in a mercuric chloride 0.1% added with two drops of Tween 20 especially for axillary and cotyledonary buds for 5 min while 0.01% of mercuric chloride was used for apical buds. The explants were cultured on 糓S and 絎PM media supplemented with BAP, KIN at different concentrations (0.2 mg/L and 3.5 mg/L respectively) and NAA (0.05 mg/L). Results: The best sprouting rate was obtained with the variety Wossro which showed a high bud break rate (26.47%) against (9.88%) for the variety Sissro. The topophysis significantly influenced the response of the buds in tissue culture. 24.48% of axillary buds were sprouted on � MS medium + 3.5 mg/L BAP+ 0.05mg/L NAA. The in vitro germination of embryos was significantly (p? 0.05) influenced by the genotype of the variety. 50.76% of buds were germinated for Wossro while 18.32% were germinated for Sissro. � MS + 0.2 mg/L KIN has significant influenced the plants growth and development. Implication: The findings will help to improve the regeneration rate and plantlets production of African bush mango tree.

Tratamiento farmacológico del estrabismo / Pharmacological treatment of strabismus

La cirugía de la musculatura extraocular ha sido el estándar de atención para tratamiento quirúrgico del estrabismo por más de un siglo. A pesar del gran desarrollo técnico de la cirugía de estrabismo en la actualidad, la utilización de microscopio quirúrgico, el diseño novedoso del instrumental quirúrgico, la calidad de la sutura no reabsorbible; los avances en equipamiento y fármacos anestésicos, la misma no está exenta de complicaciones quirúrgicas, además del tiempo de recuperación que necesita el paciente para reincorporarse a sus actividades sociales, han propiciado una búsqueda permanente del tratamiento farmacológico para el estrabismo. El objetivo de esta revisión bibliográfica es analizar las distintas alternativas farmacológicas disponibles como tratamiento del estrabismo. Para su confección se consultó textos completos y artículos en idiomas español e inglés, disponible en algunas bases de datos. Concluimos que aunque se han estudiado numerosos fármacos, la toxina botulínica que es la más conocida y utilizada mundialmente, seguida de la bupivacaína. Encontramos otros como la IGF I y II (Insuline Growing Factor), capaces de generar un efecto de reforzamiento de la actividad muscular. Y otros que "debilitan" la musculatura extraocular, incluyen la mAb35-Rubicina, BMP4 (Proteína morfogénica ósea). Se continúa su investigación en la actualidad(AU)

Extraocular musculature surgery has been the standard of care for surgical treatment of strabismus for more than a century. Despite the great technical development of strabismus surgery today, the use of a surgical microscope, the novel design of surgical instruments, the quality of the non-absorbable suture; Advances in anesthetic equipment and drugs, it is not exempt from surgical complications, in addition to the recovery time that the patient needs to return to their social activities, have led to a permanent search for pharmacological treatment for strabismus. The objective of this bibliographic review is to analyze the different pharmacological alternatives available as a treatment for strabismus. For its preparation, full texts and articles in Spanish and English languages were consulted, available in some databases. We conclude that although numerous drugs have been studied, botulinum toxin, which is the best known and used worldwide, followed by bupivacaine. We find others such as IGF I and II (Insuline Growing Factor), capable of generating an effect of reinforcing muscle activity. And others that "weaken" MOE include mAb35-Rubicin, BMP4 (Bone Morphogenic Protein). His research is continuing today(AU)

Effect of platelet-derived growth factor (PDGF-BB) and bone morphogenic protein 2 (BMP-2) transfection of rBMSCs compounded with platelet-rich plasma on adipogenic differentiation

The aim of this study was to inhibit adipogenic differentiation by transfecting two growth factors, platelet-derived growth factor (PDGF-BB) and bone morphogenic protein 2 (BMP-2), into modified rat bone marrow mesenchymal stem cells (rBMSCs) and then compounded with platelet-rich plasma (PRP). To achieve rBMSCs, the osteoporosis model of rats was established, and then the rBMSCs from the rats were isolated and identified. Co-transfection of rBMSCs with PDGF-BB-GFP and BMP-2 and detection of PDGF-BB/BMP-2 expression in transfected BMSCs was assessed by qRT-PCR and western blot, respectively. Moreover, the effect of the two growth factors transfection of rBMSCs on adipogenic differentiation was evaluated by oil red O staining and western blot, respectively. Finally, construction of the two growth factors transfection of rBMSCs compounded with PRP and detection of adipogenic differentiation were assessed by oil red O staining, CCK-8, and western blot, respectively. In vitro studies revealed that the two growth factors transfection of rBMSCs compounded with PRP promoted cell viability and inhibited adipogenic differentiation and could be promising for inhibiting adipogenic differentiation.

Effect of rhBMP-2 and Chitosan in Differentiation of Periodontal Ligament Stem Cells into an Osteoblastic Lineage

Aims: To analyze the effect of rhBMP-2 and Chitosan in differentiation of Periodontal Ligament Stem Cells (PDLC) into an osteoblastic lineage. Study Design: This study was designed as in vitro study and osteogenic biomarkers were determined in the culture supernatant. Place and Duration of Study: Laboratory of Oral Biology Faculty of Dentistry Universitas Indonesia. Jakarta 10430 Indonesia, January – September 2016. Methodology: Human periodontal ligament stem cells (PDLC) were isolated from the root of vital teeth, followed by identification of stem cells by antibody anti  STRO-1. Chitosan was used at the concentration of 0.15%.  The culture cells were divided into four groups as follow, the control group (PDLC) and treatment groups with recombinant human Bone Morphogenic protein 2 (rhBMP-2), the combination chitosan-rhBMP-2 and chitosan only. The levels of alkaline phosphatase (ALP) was determined by colorimetry and osteocalcin and collagen type I were measured using ELISA. Results: The results showed that levels of ALP tended to increase is in all groups. At day 14, the highest levels of ALP was in chitosan treated group. The concentration of collagen type 1 managed to raise is in all groups on days 14, and the highest levels Collagen type 1 occurred in RH BMP-2 and chitosan treated cells, after that decrease in all groups until day 21(p < 0.05).  Osteocalcin concentration tended to increase is in all groups, and at days 21, the highest levels in with rhBMP-2 + chitosan.   Conclusion: The rhBMP-2, chitosan, and its combination induce differentiation of periodontal ligament stem cells into the osteoblastic lineage.

Cellular Response of Anodized Titanium Surface by Poly(Lactide-co-Glycolide)/Bone Morphogenic Protein-2

BACKGROUND: The purpose of this study is to examine physical characteristics of and initial biological properties to anodized titanium treated with poly(D,L-lactide-co-glycolide) (PLG) mixed with recombinant human bone morphogenic protein-2 (rhBMP-2). METHODS: Titanium specimens were prepared in groups of four as follows: group NC was anodized under 300 V as control; group PC was anodized then dropped and dried with solution 0.02 ml PLG; group D was anodized then dropped and dried with solution 0.02 ml PLG/rhBMP-2 (3.75 µg per disc); and group E was anodized then coated with 0.02 ml PLG/rhBMP-2 (3.75 µg per disc) by electrospray. Human osteoblastic-like sarcoma cells were cultured. Cell proliferation and alkaline phosphatase (ALP) activity test were carried out. Runx-2 gene was investigated by the reverse transcription-polymerase chain reaction. Immunofluorescence outcome of osteogenic proteins was observed. RESULTS: After 3 days, there were significantly higher proliferations compared rhBMP-2 loaded titanium discs with rhBMP-2 unloaded discs. The ALPase activity on rhBMP-2 loaded titanium discs was significantly higher than in rhBMP-2 unloaded discs. The expression level of Runx2 mRNA presented the highest on the PLG/rhBMP-2-coated surface. CONCLUSION: PLG polymers mixed with rhBMP-2 might improve proliferation, differentiation and osteogenic protein formation of cells on the anodized titanium.

Does Recombinant Human Bone Morphogenic Protein 2 Affect Perioperative Blood Loss after Lumbar and Thoracic Spinal Fusion?

STUDY DESIGN: Retrospective cohort design. PURPOSE: This study aimed to determine whether recombinant human bone morphogenic protein 2 (rhBMP-2) reduces total perioperative blood loss during lumbar and thoracic fusion. OVERVIEW OF LITERATURE: Previous studies on rhBMP-2 versus iliac crest bone grafting in thoracic and lumbar fusions have yielded mixed results regarding reductions in blood loss and have largely neglected the postoperative period when analyzing total blood loss. Additionally, these studies have been limited by heterogeneity and sample size. METHODS: We analyzed the blood loss patterns of 617 consecutive adult patients undergoing lumbar and/or thoracic fusions requiring subfascial drain placement at a single institution from January 2009 to December 2016. Patients were divided into BMP and non-BMP cohorts, and a propensity score analysis was conducted to account for the differences between cohorts. RESULTS: At a per-level fused basis, the BMP group exhibited a significant reduction in the intraoperative (66.1 mL per-level fused basis; 95% confidence interval [CI], 127.9 to 4.25 mL; p=0.036) and total perioperative blood loss (100.7 mL per-level fused basis; 95% CI, 200.9 to 0.5 mL; p=0.049). However, no significant differences were observed in an analysis when not controlling for the number of levels or when examining the postoperative drain output. CONCLUSION: RhBMP-2 appears to reduce both intraoperative and total blood loss during lumbar and thoracic fusions on a per-level fused basis. This total reduction in blood loss was achieved via intraoperative effects because RhBMP-2 had no significant effect on the postoperative drain output.

Alginate encapsulation in Glycyrrhiza glabra L. with phyto-chemical profiling of root extracts of in vitro converted plants using GC-MS analysis

Objective To investigate the conversion potential of alginate encapsulated nodes of Glycyrrhiza glabra with phyto-chemical evaluation of root extract of field transferred plants. Methods The excised axenic nodal segments were encapsulated in alginate matrix planted on Murashige and Skoog (1962) medium with different supplementation and formulations of PGRs. The two year old field transferred plants were evaluated for phyto-compounds analysis using GC-MS technique. Results Varied responses were observed during the study, maximum conversion 95.83% ± 2.40% was obtained in these encapsulates when planted on MS medium containing 2.5 μM Kinetin and 0.5 μM α-Naphthalene acetic acid, which eventually developed into complete plantlets in a single step. Further, GC-MS analysis showed the presence of different phyto-compounds in the methanolic root extracts of in vitro converted plants. The results obtained revealed the presence of about 47 phyto-compounds along with various potential bioactive compounds useful for industrial and pharmaceutical purposes. Conclusions This study investigates high frequency regeneration and conversion of Glycyrrhiza glabra in a single step in short time. Also, the in vitro raised plants are analysed for bioactive compounds after field transfer, which shows the presence of numerous compounds useful for commercial and pharmacological purposes.

Alginate encapsulation in Glycyrrhiza glabra L.with phyto-chemical profiling of root extracts of in vitro converted plants using GC-MS analysis

Objective: To investigate the conversion potential of alginate encapsulated nodes of Glycyrrhiza glabra with phyto-chemical evaluation of root extract of field transferred plants. Methods: The excised axenic nodal segments were encapsulated in alginate matrix planted on Murashige and Skoog (1962) medium with different supplementation and formulations of PGRs.The two year old field transferred plants were evaluated for phyto-compounds analysis using GC-MS technique. Results: Varied responses were observed during the study, maximum conversion 95.83% ± 2.40% was obtained in these encapsulates when planted on MS medium containing 2.5 μM Kinetin and 0.5 μM α-Naphthalene acetic acid, which eventually developed into complete plantlets in a single step.Further,GC-MS analysis showed the presence of different phyto-compounds in the methanolic root extracts of in vitro con-verted plants. The results obtained revealed the presence of about 47 phyto-compounds along with various potential bioactive compounds useful for industrial and pharmaceu-tical purposes. Conclusions: This study investigates high frequency regeneration and conversion of Glycyrrhiza glabra in a single step in short time. Also, the in vitro raised plants are analysed for bioactive compounds after field transfer, which shows the presence of numerous compounds useful for commercial and pharmacological purposes.

Estudo in vitro dos efeitos da BMP-2 e do seu antagonista Noggin sobre a proliferação e migração celulares em carcinoma epidermóide de língua / In vitro study of the effects of BMP-2 and its antagonist Noggin on cell proliferation and migration in tongue squamous cell carcinoma

O carcinoma epidermóide oral (CEO) representa a neoplasia maligna mais prevalente na cavidade oral e por atingir um grande número de indivíduos, acaba se tornado um relevante problema de saúde pública. Muitos estudos demonstram alterações nos componentes da via BMP em vários tipos de tumores, como os de próstata, cólon, mama, gástricos e CEOs. É do conhecimento atual que essas proteínas podem exercer efeito pró-tumoral em estágios mais avançados do desenvolvimento neoplásico vindo a favorecer a progressão e invasão tumoral. A inibição da via de sinalização da BMP-2, através dos seus antagonistas, tem mostrado resultados positivos de ação antitumoral e que assim, o uso do Noggin pode ser um novo alvo terapêutico contra o câncer. Diante destas evidências e dos escassos trabalhos com BMP-2, Noggin e CEO, o objetivo desta pesquisa foi avaliar o efeito da BMP-2 e seu antagonista Noggin sobre a proliferação e migração celulares em culturas de células de carcinoma epidermóide de língua humana (SCC25). Foi feita a divisão em três grupos de estudo, um grupo controle, onde as células SCC25 não sofriam tratamento com substância alguma, um grupo BMP-2, no qual as células eram tratadas com 100ng/ml de BMP-2 e um grupo de células que eram tratadas com 100ng/ml de Noggin. Para o ensaio de proliferação e ciclo celular foram estabelecidos três intervalos de tempo (24, 48 e 72 horas).

A atividade proliferativa foi investigada por azul de tripan e a análise do ciclo celular através da marcação por iodeto de propídio em Citometria de fluxo. O potencial de migração/invasão das células SCC25 foi avaliado através da realização de um ensaio de invasão celular utilizando o matrigel em um intervalo de 48 horas. A curva de proliferação revelou maior crescimento celular nas células tratadas com BMP-2 no intervalo de 72 horas (p<0.05) e menor crecimento e viabilidade celular no grupo Noggin. As proteínas recombinantes favoreceram a maior porcentagem das células na fase do ciclo celular Go/G1 com diferença estatisticamente significativa no intervalo de 24 horas (p<0,05). A BMP-2 promoveu uma maior invasão das células estudadas, assim como o seu antagonista Noggin inibiu a invasão das células estudadas (p<0,05). Dessa forma, os resultados indicam que a BMP-2 favorece o fenótipo maligno, pois estimula a proliferação e invasão das células SCC25 e seu antagonista Noggin pode ser uma alternativa terapêutica pois inibiu essas características pró-tumorais. (AU)

Oral squamous cell carcinoma (OSCC) is the most prevalent malignancy in the oral cavity and reach a large number of individuals, has become an important public health problem. Studies have demonstrated changes in pathway components BMP in various types of cancers as prostate, colon, breast, gastric and OSCCs. Is the current knowledge that these proteins may exert pro-tumor effect in more advanced stages of neoplastic development coming to favor progression and invasion tumor. The inhibition of the signaling pathway BMP-2 through its antagonists, have shown positive results of antitumor activity and use of Noggin may be a novel therapeutic target for cancer. Given this evidence and the few studies with BMP-2, Noggin and OSCC, the objective of this research was to evaluate the effect of BMP-2 and its antagonist Noggin on proliferation and migration cell in line of cell cultures of human tongue squamous cell carcinoma (SCC25). The study was divided in three groups, a control group, where SCC25 cells suffered no treatment, a BMP-2 group, in which cells were treated with 100ng/ml of BMP-2 and a group of cells that were treated with 100ng/ml of Noggin.

For the proliferation assay and cell cycle were established three time intervals (24, 48 and 72 hours). Proliferative activity was investigated by trypan blue and cell cycle analysis by staining with propidium iodide flow cytometry. The potential for migration / invasion of SCC25 cells was performing by a cell invasion assay using Matrigel in a 48-hour interval. The proliferation curve showed a higher proliferation in cells treated with BMP-2 in 72 hours (p < 0.05), and lower overgrowth and cell viability in Noggin group. Recombinant proteins favored a greater percentage of cells in cell cycle phase Go/G1 with a statistically significant difference in the interval of 24 hours (p < 0.05). BMP- 2 produced a greater invasion of cells studied as well as its antagonist Noggin inhibits invasion of cells (p < 0.05). Thus, these results indicate that BMP-2 promotes malignant phenotype, dues stimulates proliferation and invasion of SCC25 cells and, its antagonist Noggin may be an alternative treatment, due to inhibit the tumor progression. (AU)

Bone Morphogenic Protein Is a Viable Adjunct for Fusion in Minimally Invasive Transforaminal Lumbar Interbody Fusion

STUDY DESIGN: Comparison of prospectively collected data of patients undergoing minimally invasive surgery transforaminal lumbar interbody fusion (MIS-TLIF) with and without recombinant human bone morphogenic protein 2 (BMP). PURPOSE: To compare the clinical, radiological outcome and complications of patients undergoing MIS-TLIF with and without BMP. OVERVIEW OF LITERATURE: BMP is an effective fusion enhancer with potential complications. Direct comparison of MIS-TLIF with and without BMP is limited to retrospective studies with short follow-up. METHODS: From June 2005 to February 2011, consecutive cases of MIS-TLIF performed by a single surgeon were included. North American Spine Society (NASS) score, Oswestry disability index (ODI), Short Form-36 (SF-36), and visual analogue score (VAS) were assessed preoperatively and at 6 and 24 months postoperatively. Fusion rates and complications were noted. RESULTS: The 252 cases comprised 104 non-BMP and 148 BMP cases. The BMP group was significantly older (mean age, 60.2 vs. 53.9; p<0.01). Preoperative scores were similar. Immediate postoperative morphine usage was significantly lower in the BMP group (12.4 mg vs. 20.1 mg, p<0.01). At 6 months, the BMP group had lower VAS back and leg pain scores (p<0.01). At 2 years, the BMP group had better leg pain scores (p<0.01), ODI (15.4 vs. 20.3, p=0.04) and NASS scores (8.8 vs. 15.8, p<0.01). Both groups showed significant clinical improvement compared to their preoperative levels. The BMP group attained a significantly higher rate of fusion at 6 months follow-up (88.4% vs. 76.8%, p=0.016) with no difference at 2 years. The non-BMP and BMP group had 12 (11.5%) and 9 (6.1%) complications and 5 (4.8%) and 2 (1.4%) reoperations, respectively. CONCLUSIONS: The use of BMP to augment fusion in MIS-TLIF is an acceptable alternative that has potential benefits of less pain in early and intermediate postoperative follow-up.

High glucose and palmitate increases bone morphogenic protein 4 expression in human endothelial cells

Here, we investigated whether hyperglycemia and/or free fatty acids (palmitate, PAL) aff ect the expression level of bone morphogenic protein 4 (BMP4), a proatherogenic marker, in endothelial cells and the potential role of BMP4 in diabetic vascular complications. To measure BMP4 expression, human umbilical vein endothelial cells (HUVECs) were exposed to high glucose concentrations and/or PAL for 24 or 72 h, and the effects of these treatments on the expression levels of adhesion molecules and reactive oxygen species (ROS) were examined. BMP4 loss-of-function status was achieved via transfection of a BMP4-specific siRNA. High glucose levels increased BMP4 expression in HUVECs in a dose-dependent manner. PAL potentiated such expression. The levels of adhesion molecules and ROS production increased upon treatment with high glucose and/or PAL, but this eff ect was negated when BMP4 was knocked down via siRNA. Signaling of BMP4, a proinflammatory and pro-atherogenic cytokine marker, was increased by hyperglycemia and PAL. BMP4 induced the expression of infl ammatory adhesion molecules and ROS production. Our work suggests that BMP4 plays a role in atherogenesis induced by high glucose levels and/or PAL.

Enhancement of Osseointegration of Artificial Ligament by Nano-Hydroxyapatite and Bone Morphogenic Protein-2 into the Rabbit Femur

The MTT assay showed that the cell proliferation on hydroxyapatite (HAp) and HAp/bone morphogenic protein (BMP) coated group was better than the control and BMP coated groups at 5 days. And after 7 days of culture, the mRNA expression levels of type I collagen, osteonectin, osteopontin, bonesialoprotein, BMP-2, alkaline phosphatase (ALP) and Runx-2 in the HAp/BMP coated group were significantly higher than the other groups. Also, in this group showed the most significant induction of osteogenic gene expression compared to mesenchymal stem cells (MSCs) grown on the other groups. In addition, the cells in the HAp/BMP coated group delivered higher levels of ALP than the other three groups. Also, silk scaffolds were implanted as artificial ligaments in knees of rabbits, and they were harvested 1 and 3 months after implantation. On gross examination, HE staining showed that new bone tissue formation was more observed in the HAp/BMP coated group 3 weeks postoperatively. And masson staining showed that in the HAp/BMP coated group, the silk fibers were encircled by osteoblast, chondrocyte, and collagen. Furthermore, the analysis showed that the width of the graft-bone interface in the HAp and HAp/BMP coated group was narrower than that in the other two groups 3 weeks postoperatively. So, it is concluded that BMP incorporated HAp coated silk scaffold can be enhanced osseointegration and osteogenesis in bone tunnel. As a result, these experimental designs have been demonstrated to be effective in the acceleration of graft-to-bone healing by increasing new bone or fibrocartilage formation at the interface between graft and bone.

Capim-piatã adubado com fontes de fósforo de diferente solubilidade em água / Piata palisadegrass fertilized with different sources of phosphorus for solubility in water

The development of piatã grass fertilized with phosphorus sources with different solubilities in water was evaluated. The experiment was developed in the unit of Aquidauana of UEMS, MS, from February to August, 2012. Four treatments were evaluated: control (without phosphorus fertilization), slow solubility, fast solubility (readily available), and mixed solubility sources of phosphorus (FH Pasture(r)). It was adopted a randomized blocks design with four blocks and four replicates in a total of 16 pastures (0.5 ha each one). Measurements were performed weekly to evaluate structural characteristics and morphogenic variables of the plants. Data were submitted to a variance analysis according to the model. When necessary, the means of treatments with different phosphorus fertilizer sources were compared with the control using the Dunnett test and with other treatments by using student's t test. The significance level of 5% was adopted in all analysis. No significant effect was observed (P 0.05) in the evaluated traits in the first year of the experiment.

Estudo in vitro dos efeitos da BMP-2 e do seu antagonista Noggin sobre a proliferação e migração celulares em carcinoma epidermóide de língua / In vitro study of the effects of BMP-2 and its antagonist Noggin on cell proliferation and migration in squamous carcinoma of the tongue

O carcinoma epidermóide oral (CEO) representa a neoplasia maligna mais prevalente na cavidade oral e por atingir um grande número de indivíduos, acaba se tornado um relevante problema de saúde pública. Muitos estudos demonstram alterações nos componentes da via BMP em vários tipos de tumores, como os de próstata, cólon, mama, gástricos e CEOs. É do conhecimento atual que essas proteínas podem exercer efeito pró-tumoral em estágios mais avançados do desenvolvimento neoplásico vindo a favorecer a progressão e invasão tumoral. A inibição da via de sinalização da BMP-2, através dos seus antagonistas, tem mostrado resultados positivos de ação antitumoral e que assim, o uso do Noggin pode ser um novo alvo terapêutico contra o câncer. Diante destas evidências e dos escassos trabalhos com BMP-2, Noggin e CEO, o objetivo desta pesquisa foi avaliar o efeito da BMP-2 e seu antagonista Noggin sobre a proliferação e migração celulares em culturas de células de carcinoma epidermóide de língua humana (SCC25). Foi feita a divisão em três grupos de estudo, um grupo controle, onde as células SCC25 não sofriam tratamento com substância alguma, um grupo BMP-2, no qual as células eram tratadas com 100ng/ml de BMP-2 e um grupo de células que eram tratadas com 100ng/ml de Noggin. Para o ensaio de proliferação e ciclo celular foram estabelecidos três intervalos de tempo (24, 48 e 72 horas). A atividade proliferativa foi investigada por azul de tripan e a análise do ciclo celular através da marcação por iodeto de propídio em Citometria de fluxo. O potencial de migração/invasão das células SCC25 foi avaliado através da realização de um ensaio de invasão celular utilizando o matrigel em um intervalo de 48 horas. A curva de proliferação revelou maior crescimento celular nas células tratadas com BMP-2 no intervalo de 72 horas (p<0.05) e menor crecimento e viabilidade celular no grupo Noggin. As proteínas recombinantes favoreceram a maior porcentagem das células na fase do ciclo celular Go/G1 com diferença estatisticamente significativa no intervalo de 24 horas (p<0,05). A BMP-2 promoveu uma maior invasão das células estudadas, assim como o seu antagonista Noggin inibiu a invasão das células estudadas (p<0,05). Dessa forma, os resultados indicam que a BMP-2 favorece o fenótipo maligno, pois estimula a proliferação e invasão das células SCC25 e seu antagonista Noggin pode ser uma alternativa terapêutica pois inibiu essas características pró-tumorais. (AU)

Oral squamous cell carcinoma (OSCC) is the most prevalent malignancy in the oral cavity and reach a large number of individuals, has become an important public health problem. Studies have demonstrated changes in pathway components BMP in various types of cancers as prostate, colon, breast, gastric and OSCCs. Is the current knowledge that these proteins may exert pro-tumor effect in more advanced stages of neoplastic development coming to favor progression and invasion tumor. The inhibition of the signaling pathway BMP-2 through its antagonists, have shown positive results of antitumor activity and use of Noggin may be a novel therapeutic target for cancer. Given this evidence and the few studies with BMP-2, Noggin and OSCC, the objective of this research was to evaluate the effect of BMP-2 and its antagonist Noggin on proliferation and migration cell in line of cell cultures of human tongue squamous cell carcinoma (SCC25). The study was divided in three groups, a control group, where SCC25 cells suffered no treatment, a BMP-2 group, in which cells were treated with 100ng/ml of BMP2 and a group of cells that were treated with 100ng/ml of Noggin. For the proliferation assay and cell cycle were established three time intervals (24, 48 and 72 hours). Proliferative activity was investigated by trypan blue and cell cycle analysis by staining with propidium iodide flow cytometry. The potential for migration / invasion of SCC25 cells was performing by a cell invasion assay using Matrigel in a 48-hour interval. The proliferation curve showed a higher proliferation in cells treated with BMP-2 in 72 hours (p < 0.05), and lower overgrowth and cell viability in Noggin group. Recombinant proteins favored a greater percentage of cells in cell cycle phase Go/G1 with a statistically significant difference in the interval of 24 hours (p < 0.05). BMP- 2 produced a greater invasion of cells studied as well as its antagonist Noggin inhibits invasion of cells (p < 0.05). Thus, these results indicate that BMP-2 promotes malignant phenotype, dues stimulates proliferation and invasion of SCC25 cells and, its antagonist Noggin may be an alternative treatment, due to inhibit the tumor progression. (AU)

Morphogenesis and production of tanzânia, mombaça and mulato grasses under nitrogen fertilization / Morfogênese e produção dos capins tanzânia, mombaça e mulato sob adubação nitrogenada

A field trial was conducted designed in a completely randomized block in a 4 x 3 factorial arrangement to evaluate the application of nitrogen doses (N) (0, 40, 80 and 160 kg/ha) on the morphogenical characteristics and dry matter partition of three forage grasses (Panicum maximum cvs. Mombasa and Tanzania and Brachiaria sp. Hybrid Mulato). The leaf appearance (LAR, leaf/day) and stretching (LER; mm/day) rates, the number of green leaves per tiller (NLT) and the average weight of tillers (MTW; g) presented s positive linear response to the N dose while the phyllochron (Phil; day/leaves) showed a negative linear response. The highest LER, IAL and final leaf length (FLL; cm) occurred in the Mombaça and Tanzania grasses, while the highest LAR occurred in the Mulato grass. There was a negative quadratic effect of the N dose on the stem elongation rate (SER; mm/day) and LF. The Mombaça and Tanzania grasses presented the highest SER; however, in just two forages. The production of total dry matter (TDM; kg/ha), leaves (LDM; kg/ha) and stems (SDM; kg/ha) increased linearly and quadratically with the N dose, respectively, for the Mombaça and Tanzania grasses. There was a high positive correlation among DM, LDM and SDM and the Mombaça grass MTW. The dry matter production and morphogenic characteristics were influenced by the nitrogen fertilization as a result of the substantial increase in the flow of tissues stimulated by fertilization, proving the importance of N for forage biomass accumulation.

O experimento foi conduzido sob o delineamento em blocos completos ao acaso em esquema fatorial 4x3 para avaliar a aplicação de doses de nitrogênio (N) (0, 40, 80 e 160 kg/ha) sobre as características morfogênicas e a partição de matéria seca (MS) de três gramíneas forrageiras (Panicum maximum cvs. Mombaça e Tanzânia e Brachiaria sp. híbrida Mulato). As taxas de aparecimento (TapF; folhas/dia) e alongamento de folhas (TalF; mm/folha), o número de folhas verdes por perfilho (NFVP) e o peso médio de perfilhos (PMP; g) apresentaram resposta linear positiva às doses de N enquanto o filocrono (Fil; dias/folha) apresentou resposta linear negativa. Os capins Tanzânia e Mombaça apresentaram maiores TalF, Fil e comprimento final de folhas (CFF; cm) enquanto o capim Mulato apresentou maior TapF. As produções de matéria seca total (PMST; kg/ha) de folhas (PMSF; kg/ha) e de colmos (PMSC; kg/ha) aumentaram de forma quadrática e linear com as doses de N, respectivamente, para os capins Mombaça e Tanzânia. Houve alta correlação positiva entre as PMST, PMSF e PMSC e o PMP da forrageira Mombaça. A produção de matéria seca e as características morfogênicas foram influenciadas pela adubação nitrogenada, como resultado do aumento substancial do fluxo de tecidos estimulado pela adubação, o que comprova a importância do N para o acúmulo de biomassa da forragem.

Effect of mechanical centrifugal force on Runx-2 mRNA in osteoblasts’ bone morphogenetic protein signal pathway / 中国组织工程研究

BACKGROUND:Mechanical strain certainly has an effect on physiological activities of osteoblasts. Runx-2 is a target of bone morphogenic protein signal and is an important factor for regulation of osteoblastic differentiation. Bone morphogenic protein signal transduction pathway is involved in physiological response of osteoblast to stimulation of mechanical centrifugal force. OBJECTIVE:To observe the effect of mechanical centrifugal force on bone morphogenetic protein signal pathway under different time period and speed. METHODS:MC3T1-E1 cells were pre-treated in DMEM medium containing 10%fetal bovine serum for 24 hours, and then divided into control group, 90 r/min group, 180 r/min group and 250 r/min group. Each group was then subdivided into 6 hours, 12 hours and 24 hours centrifugation subgroups. Experiments were repeated for three times for different centrifugal speed and different time period. Except centrifugation, the control group was under the same environment. Total RNA was extracted and reversely transcribed into cDNA. Runx-2 gene expression was determined by real time fluorescent quantitative PCR. RESULTS AND CONCLUSION:The expression of Runx-2 mRNA was increased with extension of time, showing a positive correlation between the two. The mRNA expression at 180 r/min was significantly higher than that at 90 r/min and 250 r/min (P<0.01);at 90 r/min and 180 r/min, the Runx-2 mRNA expression was higher than that in the control group (P=0.039), both of them showed significant difference along with the time. The difference of centrifugal force speed and duration is associated with different physiological response of osteoblasts in bone morphogenic protein signal pathway, which plays an important role in mechanical signal transmission and cascade reaction.

Effects of apigen on repairing articular cartilage defects by BMP-7 cells in rabbits / 中华临床营养杂志

Objective To study the effect of apigen on articular cartilage repair involving chondrocytes transfected with bone morphogenic protein-7 (BMP-7).Methods lnterleukin (IL-8) and soluble intercellular adhesion molecule-1 (sICAM-1) were induced by IL-1 β in rabbit chondrocytes.After apigen at different concentrations (10 μmol/L,25 μmol/L,and 50 μmol/L) was added into the culture system,the effect of apigen on IL8 and sICAM-1 production was observed using enzyme linked immunosorbent assay.Then chondrocytes were seeded on improved matrigel gel bracket and culturedin vitro to construct the compound,which was then transplanted into the rabbit model of articular cartilage defection.The rabbits were randomly divided into sham group (n =4),trans-BMP-7 group (n =4),and trans-BMP-7 + apigen group (n =4).Histological observation was conducted and Wakitani score calculated after 5 weeks.Results The concentrations of IL-8 and sICAM-1 in the chondrocytes supernatant in vitro significantly decreased after apigen treatment at 10 μmol/L,25 μmol/L,and 50 μmol/L [(6803.63 ±162.31) ng/g,(6005.74 ±201.49) ng/g,and (5202.34 ±271.67) ng/gvs.(10011.84±239.29) ng/g ; P =0.00].Five weeks after the cartilage cells on matrigel gel bracket were transplanted into rabbit models,the Wakitani scores of the trans-BMP-7 group and the trans-BMP-7 + apigen group were significantly lower than that of the sham group [(3.68 ± 0.86) vs.(8.25 ± 0.90),P =0.00 ; (3.21 ± 0.78) vs.(8.25 ±=0.90),P =0.00].In addition,no inflammatory reaction was noted during the repair in the trans-BMP-7 + apigen group.Conclusion Apigen can promote the construction of compound and repair of articular cartilage defects by trans-BMP-7 chondrocyte.

The evaluation of the correlation between histomorphometric analysis and micro-computed tomography analysis in AdBMP-2 induced bone regeneration in rat calvarial defects

PURPOSE: Micro-computed tomography (micro-CT) has been widely used in the evaluation of regenerated bone tissue but the reliability of micro-CT has not yet been established. This study evaluated the correlation between histomorphometric analysis and micro-CT analysis in performing new bone formation measurement. METHODS: Critical-size calvarial defects were created using a 8 mm trephine bur in a total of 24 Sprague-Dawley rats, and collagen gel mixed with autogenous rat bone marrow stromal cells (BMSCs) or autogenous rat BMSCs transduced by adenovirus containing bone morphogenic protein-2 (BMP-2) genes was loaded into the defect site. In the control group, collagen gel alone was loaded into the defect. After 2 and 4 weeks, the animals were euthanized and calvaria containing defects were harvested. Micro-CT analysis and histomorphometric analysis of each sample were accomplished and the statistical evaluation about the correlation between both analyses was performed. RESULTS: New bone formation of the BMP-2 group was greater than that of the other groups at 2 and 4 weeks in both histomorphometric analysis and micro-CT analysis (P=0.026, P=0.034). Histomorphometric analysis of representative sections showed similar results to histomorphometric analysis with a mean value of 3 sections. Measurement of new bone formation was highly correlated between histomorphometric analysis and micro-CT analysis, especially at the low lower threshold level at 2 weeks (adjusted r2=0.907, P<0.001). New bone formation of the BMP-2 group analyzed by micro-CT tended to decline sharply with an increasing lower threshold level, and it was statistically significant (P<0.001). CONCLUSIONS: Both histomorphometric analysis and micro-CT analysis were valid methods for measurement of the new bone in rat calvarial defects and the ability to detect the new bone in micro-CT analysis was highly influenced by the threshold level in the BMP-2 group at early stage.

Guided Bone Regeneration Using a Putty-type Demineralized Bone Matrix: Case Report

Effects of Polycaprolactone-Tricalcium Phosphate, Recombinant Human Bone Morphogenetic Protein-2 and Dog Mesenchymal Stem Cells on Bone Formation: Pilot Study in Dogs

PURPOSE: The aim of this study was to evaluate the survival, proliferation, and bone formation of dog mesenchymal stem cells (dMSCs) in the graft material by using Polycaprolactone-tricalcium phosphate (PCL-TCP), auto-fibrin glue (AFG), recombinant human bone morphogenetic protein-2 (rhBMP-2), and dMSCs after a transplantation to the scapula of adult beagle dogs. MATERIALS AND METHODS: The subjects were two beagle dogs. Total dose of rhBMP-2 on each block was 10 microg with 50 microg/mg concentration. The cortical bone of the scapula of the dog was removed which was the same size of PCL-TCP block (Osteopore International Pte, Singapore; 5.0x5.0x8.0 mm in size), and the following graft material then was fixed with orthodontic mini-implant, Dual-top(R) (Titanium alloy, Jeil Co. Seoul, Korea). Four experimental groups were prepared for this study, Group 1: PCL-TCP + aFG; Group 2: PCL-TCP + aFG + dMSCs; Group 3: PCL-TCP + aFG + dMSCs + rhBMP-2; Group 4: PCL-TCP + aFG + dMSCs + rhBMP-2 + PCL membrane. The survival or proliferation of dMSCs cells was identified with an extracted tissue through a fluorescence microscope, H-E staining and Von-Kossa staining in two weeks and four weeks after the transplantation. RESULTS: The survival and proliferation of dMSCs were identified through a fluorescence microscope from both Group 1 and Group 2 in two weeks and four weeks after the transplantation. Histological observation also found that the injected cells were proliferating well in the G2, G3, and G4 scaffolds. CONCLUSION: This study concluded that bone ingrowth occurred in PCL-TCP scaffold which was transplanted with rhBMP-2, and MSCs did not affect bone growth. More sufficient healing time would be needed to recognize effects of dMSCs on bone formation.