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.

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)

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.

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.

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)

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.

The BMPs expression and histomorphometric study of beta-TCP /rhBMP-2 Grafting on the rabbit cranial bone defects

OBJECTIVE: The Purpose of the study was to investigate the bone morphogenic protein expression of rhBMP-2(recombinant human bone morphogenic protein-2) as singnaling molecule and beta-TCP(Tricalcium phosphate) as a bone substitute and carrier medium of rhBMP-2. MATERIALS AND METHODS: 16 rabbits divided into 2 group of each 8 rabbit. Two standardized bone defect, round bilateral defect was made in the cranium of the 8 rabbit of first group, and was grafted with 150~500micrometer diameter beta-TCP 0.25g in one side, which was soaked with rhBMP-2, and autogenous bone was grafted on another side as a positive control. Second group of 8 rabbit, only beta-TCP was grafted with same size and same manner. After 2, 4, 8, and 12 weeks, specimen was taken for microscopic immunohiostochemical and histomorphometric analysis. RESULT: Grafting beta-TCP with rhBMP show the early formation of the bone regenerative factor (BMP-4) and more quantity of new bone formation than only use of beta-TCP (8,12 week), even show less new bone formation than autogenous bone. CONCLUSION: The experimental study result that beta-TCP graft combination with rhBMP-2 as a delivery system is an effective with osteoinductive capacity and biodegradable properties, so that provide clinical availibility of composite use in reconstruction of bony defect.

Gene therapy of firearm femur defect by using BMP-4/TGF-β1 in rabbits / 中华创伤杂志

Objective To observe effects of BMP-4 gene combined with TGF-β1 in repairing rabbit femur defect resulted from firearm wound. Methods The femur defect was made with firearm steel ball. Plasmid encoded BMP-4 gene identified in vitro and TGF-β1 were injected into the tissue of upper, lower and central parts of the defects at the second week after wound. The mRNA and protein expressions of BMP-4 in vivo were detected by real time-polymerase chain reaction (RT-PCR) and Western blot. Activity of alkaline phosphatase (ALP) and calcium content were measured for evaluating osteogenetic ability. The process and quality of osteogenesis were determined by pathological and X-ray examinations. Results mRNA and protein of BMP-4 could continually express for six weeks in vivo after injection. Activity of ALP in the experimental group was increased to ( 13.17 ±0.51 ) U/100 ml at the 8th week, which was significantly higher than (8.77 ± 0.44) U/100 ml in the control group, indicating that osteogenetic ability was markedly enhanced, which accorded with determination of calcium content. Pathological observation and X-ray proved prominent improvement of osteogenesis, with a shorter time and better quality, in the experimental group. Conclusion BMP-4/TGF-β1 can promote repair of firearm femur defect.

The Healing Effectof Bone Morphogenic Protein with Fibrin Glue on an Injury of the Tendon-Bone Junction / 대한정형외과학회잡지

Purpose: The author hypothesizes that exogenously injected BMP, which is mixed with fibrin glue, can accelerate the healing of a bone-tendon junction injury and increase its holding strength during the early regeneration period. Materials and Methods: A direct injury model of the bone-tendon junction was made using the Achilles tendon-calcaneus bone of 54 rabbits: and the transected Achilles tendon was repaired to its original insertion site using the Krackow method. In Group 1, no additional manipulation was performed. In Group 2, only fibrin glue was injected into the junction between the Achilles tendon and the calcaneus in order to exclude the effect of the fibrin glue. In Group 3, BMP-2 incorporated into the fibrin glue was injected into the junction. The results were evaluated by histological analysis and biomechanical tests at 2, 4, and 8 weeks after surgery. The Kruskal-Wallis test was used for a statistical evaluation. Results: Histological analysis revealed the early appearance of fibrocartilage at 2 weeks in Group 3: the area of the fibrocartilage expanded with time. The biomechanical tests showed significant differences in the maximum stress between Groups 1 and 3, and between Groups 2 and 3, at 2, 4, and 8 weeks. 74.4% of the normal maximum stress was recovered at 8 weeks in Group 3. Conclusion: The combined use of BMP-2 and the fibrin glue can accelerate the healing of an injury of the bone-tendon junction.

Experimental study of expression of BMP-2 and VEGFmRNA in local femoral head of nontraumatic osteonecrosis of femoral head / 中国矫形外科杂志

[Objective]To explore the change and its significance of the expression of vascular endothelial growth factor(VEGF) messenger RNA and bone morphogenic protein-2(BMP-2) of the local femoral head in the nontraumatic osteonecrosis of femoral head(NONFH).[Method]Sixteen samples of femoral heads of NONFH were collected as the experimental group and fresh 10 samples of femoral heads of femoral neck fracture as control group,overall examples were collected from total articular replacement arthroplasty.The samples were splitted in coronal plane and get one bone block from necrosis area and another one from healthy area,make them into microtome section after the process of immobility and decalcification.Their pathological change was observed by using optical microscope and electron microscope and detect the expression of BMP-2 and VEGFmRNA in femoral head through making use of immunohistochemistry and in-situ hybridization technique.[Result]The organization structure of experimental group was disorganized,cracked and the bone trabecula was rarefactive and non-intact and there was a great number of empty lacuna in bone trabecula.While there was the reverse situation in the control group.The intensity and area of positive expression of BMP-2 and VEGFmRNA in femoral head of the experimental group were lower than that of control group obviously.The result showed statistical significance(P

Cell Specific Non-Viral Vector Enhances Expression of BMP-2 Gene in Human Dermal Fibroblasts / 上海第二医科大学学报

Objective To construct the fibroblast-specific non-viral vector pcDNA3-CEP-BMP-2 containing collagen 1A2 enhancer and promoter,and to validate the enhancement of BMP-2 expression in the human dermal fibroblasts by this vector,compared with the routine non-viral BMP2 vector. Methods The sequences for collagen 1A2 enhancer and promotor,and BMP-2 gene were ligated into the pcDNA3 plasmids.The plasmids were transfected into human skin fibroblasts and vein endothelial cells by means of cationic liposomes.The expressions of the plasmids in these two kinds of cells were detected by RT-PCR.The osteogenic phonotypes of fibroblasts were determined.(Results)pcDNA3-CEP-BMP-2,which contained collagen 1A2 enhancer and promoter could enhance the BMP-2 expression in the fibroblasts but not in vein endothelial cells.Osteogenetic phenotypes were more obvious in the fibroblasts transfected with pcDNA3-CEP-BMP-2 than in pcDNA3-BMP-2-transfected ones. Conclusion Collagen 1A2 enhancer and promoter can enhance BMP2 expression in fibroblasts.

Effect of Chitosan, betaig-h3 and Human Bone Morphogenic Protein-4 on Early Bony Consolidation in Distraction Osteogenesis of the Canine Mandible

Sixteen dogs were used to study the effect of bone morphogenic protein(BMP-4), betaig-h3 and chitosan during the early bony consolidation stage in the distracted zones of mandibles. The lateral surface of the mandibular body was exposed in the subperiosteal plane and vertical osteotomy was carried out on the mandibular body. An external distraction device was applied to the mandibular body about 1 cm apart from the osteotomy line. Mandibular distraction was started 5 days after the mandibular osteotomy at a rate of 2 mm per day for a total of 10 mm distraction for 5 days. The experimental group was divided into 4 groups: control group, BMP-4 group, betaig-h3 group and chitosan group depending on the injected material into the distracted area. Four dogs were allocated to each group. On the day of completion of distraction, 0.5 ml of BMP-4, 0.5 ml of betaig-h3, 0.5 ml of 5% chitosan solution was injected respectively into the distracted area of each group with the same amount of tripolyphosphate in dual syringe for solidification of the injected solution. In the control group, 1 ml of tripolyphosphate was injected into the distracted area. After injection of the study materials, the distraction device was left in place for 4 or 7 weeks to allow bony consolidation. Radiographs were taken weekly. Two dogs in each group, a total of eight dogs, were sacrified in 4 weeks, and another eight dogs in 7 weeks after completion of distraction. Bone specimens of the distracted mandibles were taken for histologic examination. The mineral density of the distracted bone was measured during the radiological procedures and analysed by the computer. In the radiographs of the distracted areas of the mandibles, the control group has shown a mostly radiolucent zone but the other groups have shown the radiodense zones with various width of central radiolucent zones. The central radiolucent zone became narrower in time and vertical thickness of the radiodense zone was about twice thicker in 7 weeks than that of 4 weeks after finishing bone distraction. BMP-4 group showed the thickest radiodense zone and the chitosan group shows the thinnest radiodense zone. The mineral density of bone was highest in the BMP-4 group and lowest in the control group. In the histological findings of the distracted areas of mandibles, the control group showed whole fibrous tissue but the other groups showed new woven bones with central narrow fibrous interzone. The degree of new bone formation was most remarkable in the BMP-4 group and was least remarkable in the chitosan group. In conclusion, there was an active formation of a new bone in the distracted area of the mandible by injection of BMP-4, betaig-h3 and chitosan. The new bone formation was most remarkable in the BMP-4 group followed by betaig-h3, chitosan and control group. These findings suggest that BMP-4 is clinically worth using for early bony consolidation in the distraction osteogenesis.