377 research outputs found
Cellular responses to deproteinized bovine bone mineral biofunctionalized with bone-conditioned medium.
OBJECTIVES
The aim of the study was to investigate whether the osteoinductive properties of bone-conditioned medium (BCM) harvested from cortical bone chips within a clinically relevant short-term period can enhance the biologic characteristics of deproteinized bovine bone mineral (DBBM) in vitro.
MATERIALS AND METHODS
To assess the biofunctionalization of DBBM, the adhesive, proliferative, and differentiation properties of mesenchymal stromal ST2, pre-osteoblastic MC3T3-E1, and primary bone-derived cells grown on BCM-coated DBBM were examined by crystal violet staining of adherent cells, BrdU ELISA, and qRT-PCR, respectively.
RESULTS
BCM extracted within 20 min or 24 h in either Ringer's solution (BCM-RS) or RS mixed with autologous serum (BCM-RS + S) increased the adhesive properties of all three cell types seeded on DBBM. The 20-min BCM-RS preparation appeared more potent than the 24-h preparation. BCM-RS made within 20 min or 24 h had strong pro-proliferative effects on all cell types grown on DBBM. RS + S alone exhibited a considerable pro-proliferative effect, suggesting an impact of the serum on cellular growth. DBBM coated with BCM-RS or BCM-RS + S, made within 20 min or 24 h each, caused a significant induction of osteogenic differentiation marker expression with a higher potency of the BCM-RS + S. Finally, a strong additive effect of fresh bone chips combined with BCM-coated DBBM on the osteogenic differentiation of the three cell types was observed.
CONCLUSIONS
Altogether, the data strongly support the biofunctionalization of DBBM with BCM extracted within a clinically relevant time window of 20 min.
CLINICAL RELEVANCE
Pre-activation of non-osteoinductive biomaterials with BCM, prepared from autologous bone chips during a guided bone regeneration (GBR) procedure, bears the potential of an optimal treatment modality for bone defects in daily practice
Osteoblast proliferation and differentiation on a barrier membrane in combination with BMP2 and TGFβ1
Objectives: Bioresorbable collagen membranes are routinely utilized in guided bone regeneration to selectively direct the growth and repopulation of bone cells in areas of insufficient volume. However, the exact nature by which alveolar osteoblasts react to barrier membranes as well as the effects following the addition of growth factors to the membranes are still poorly understood. The objective of the present study was therefore to investigate the effect of a bioresorbable collagen membrane soak-loaded in growth factors bone morphogenetic protein 2 (BMP2) or transforming growth factor β1 (TGFβ1) on osteoblast adhesion, proliferation, and differentiation. Material and methods: Prior to experimental seeding, membranes were soaked in either BMP2 or TGFβ1 at a concentration of 10ng/ml for 5min. Results: Human osteoblasts adhered to all soak-loaded membranes as assessed by scanning electron microscopy. Growth factors BMP2 and TGFβ1 increased osteoblast proliferation at 3 or 5days post-seeding when compared to control collagen membranes. Analysis of real-time PCR revealed that administration of BMP2 increased osteoblast differentiation markers such as osterix, collagen I, and osteocalcin. BMP2 also increased mineralization of primary osteoblasts as demonstrated by alizarin red staining when compared to control and TGFβ1 soak-loaded membranes. Conclusion: The combination of a collagen barrier membrane with growth factors TGFβ1 and BMP2 significantly influenced adhesion, proliferation, and differentiation of primary human osteoblasts. Clinical relevance: The described in vitro effects following the combination of collagen barrier membranes with growth factors TGFβ1 and BMP2 provide further biologic support for the clinical application of this treatment strategy in guided bone regeneration procedure
Gene array of primary human osteoblasts exposed to enamel matrix derivative in combination with a natural bone mineral
Objectives: The application of an enamel matrix derivative (EMD) for regenerative periodontal surgery has been shown to promote formation of new cementum, periodontal ligament, and alveolar bone. In intrabony defects with a complicated anatomy, the combination of EMD with various bone grafting materials has resulted in additional clinical improvements, but the initial cellular response of osteoblasts coming in contact with these particles have not yet been fully elucidated. The objective of the present study was to evaluate the in vitro effects of EMD combined with a natural bone mineral (NBM) on a wide variety of genes, cytokines, and transcription factors and extracellular matrix proteins on primary human osteoblasts. Material and methods: Primary human osteoblasts were seeded on NBM particles pre-coated with versus without EMD and analyzed for gene differences using a human osteogenesis gene super-array (Applied Biosystems). Osteoblast-related genes include those transcribed during bone mineralization, ossification, bone metabolism, cell growth and differentiation, as well as gene products representing extracellular matrix molecules, transcription factors, and cell adhesion molecules. Results: EMD promoted gene expression of various osteoblast differentiation markers including a number of collagen types and isoforms, SMAD intracellular proteins, osteopontin, cadherin, alkaline phosphatase, and bone sialoprotein. EMD also upregulated a variety of growth factors including bone morphogenetic proteins, vascular endothelial growth factors, insulin-like growth factor, transforming growth factor, and their associated receptor proteins. Conclusion: The results from the present study demonstrate that EMD is capable of activating a wide variety of genes, growth factors, and cytokines when pre-coated onto NBM particles. Clinical relevance: The described in vitro effects of EMD on human primary osteoblasts provide further biologic support for the clinical application of a combination of EMD with NBM particles in periodontal and oral regenerative surger
Complications and treatment errors in implant positioning in the aesthetic zone: Diagnosis and possible solutions.
Incorrect implant positioning can lead to functional and aesthetic compromise. Implant positioning errors can occur in three dimensions: mesiodistal, corono-apical, and orofacial. Treatment solutions to manage adverse outcomes through positioning errors require an understanding of the underlying conditions and of those factors that may have led to the error being committed in the first place. These types of complications usually occur because of human factors. If errors do occur with adverse aesthetic outcomes, they are difficult and sometimes impossible to correct. Connective tissue grafts to reverse recession defects are only feasible in defined situations. The option to remove and replace the implant may be the only recourse, provided the removal process does not further compromise the site. Error in judgment by the clinician
Role of Gastric Colonization in Nosocomial Infections and Endotoxemia: A Prospective Study in Neurosurgical Patients on Mechanical Ventilation
The role of gastric microbial colonization in nosocomial infections and endotoxemia was investigated prospectively in 40 neurosurgical patients requiring mechanical ventilation for >48 h. Each was studied up to 7 d. Swabs from the nose and oropharynx were cultured at admission, and aspirates from the stomach and trachea were cultured daily until enteral alimentation was started. Patients were evaluated every second day for endotoxemia and coagulation activation. Of 153 gastric aspirates, 66.7% contained microorganisms at a mean quantity of 107 cfu/ml, Nosocomial pneumonia occurred in 15 patients, septicemia in 5, and meningitis in 1. The stomach was the evident source of infection in only 1 patient with pneumonia. Of 140 plasma samples, 12 (8.6%) from 10 patients showed detectable endotoxin levels, but there was no association between endotoxemia or coagulation activation and the presence of microorganisms in the stomach. The stomach was not an important source for nosocomial infections or endotoxemia, even in patients with high gastric p
Bone grafting material in combination with Osteogain for bone repair: a rat histomorphometric study.
OBJECTIVES
Enamel matrix derivative (EMD) has been successfully used for the regeneration of periodontal tissues including new cementum, periodontal ligament, and alveolar bone. Combination of EMD with bone grafting materials has however generated variable clinical results. Recently, we have demonstrated that a new formulation of EMD in a liquid carrier system (Osteogain®) has improved physicochemical properties for the adsorption of EMD to a bone grafting material. The aim of the present study was to investigate the regenerative potential of Osteogain®, in combination with a bone graft, on new bone formation in a rat femur defect model.
MATERIALS AND METHODS
Fifty-four critically sized femur defects (3 mm in diameter) were created bilaterally in 27 rats and treated following the group allocation: (1) drilled unfilled control, (2) a natural bone mineral (NBM), and (3) NBM + Osteogain®. All defects were histologically analyzed at 2, 4, and 8 weeks after surgical intervention. Micro-CT analysis, hematoxylin and eosin (H&E) staining, and Safranin O staining were performed to quantify new bone formation.
RESULTS
Significantly more new bone formation was observed in defects treated with NBM + Osteogain® at both 4 and 8 weeks when compared to NBM alone and the control unfilled defects (P < 0.05). Histologically, the formation of more mature mineralized bone with the presence of osteocytes were found more commonly in defects treated with Osteogain® + NBM at 8 weeks post-healing when compared to NBM alone.
CONCLUSIONS
The present study demonstrate that Osteogain® in combination with a bone grafting material improves the speed and quality of new bone formation in rat osseous defects.
CLINICAL RELEVANCE
Future clinical research are now warranted to fully characterize the benefits of Osteogain®, a new formulation of enamel matrix proteins delivered in liquid formation when used in combination with a bone grafting material
Gene array of PDL cells exposed to Osteogain in combination with a bone grafting material.
OBJECTIVES
The aim of the present study was to investigate the effects of Osteogain, a new formulation of enamel matrix derivative (EMD) in combination with a grafting material on a wide variety of genes for cytokines, transcription factors and extracellular matrix proteins involved in osteoblast differentiation.
MATERIALS AND METHODS
Primary human periodontal ligament (PDL) cells were seeded on natural bone mineral (NBM) particles coated with Osteogain for 24 h and analyzed for regulated gene expression using a human osteogenesis gene super-array kit. Osteoblast-related genes include those transcribed during bone mineralization, ossification, bone metabolism, cell growth and differentiation as well as gene products representing extracellular matrix molecules, transcription factors and cell adhesion molecules.
RESULTS
Osteogain significantly upregulated the expression of over 20 of the 100 genes examined including bone morphogenetic protein 2 (BMP2), TGFβ1, fibroblast growth factor (FGF), epidermal growth factor (EGF) and platelet-derived growth factor (PDGF) as well as some of their associated receptors. Osteogain also promoted gene expression of a number of osteoblast differentiation markers including collagen1α2 and alkaline phosphatase as well as cell adhesion molecules including fibronectin and a variety of integrin binding proteins. Interestingly, Osteogain promoted calcitonin receptor 55-fold and also promoted annexin A5 gene expression over 12-fold.
CONCLUSION
The present study demonstrates that Osteogain is capable of either upregulating or downregulating the expression of a wide variety of genes including those for growth factors and cytokines when combined with a bone grafting material.
CLINICAL RELEVANCE
The results from the present study demonstrate the large and potent effect of addition of Osteogain in combination to a bone grafting material over a wide variety of genes supporting osteogenesis
Sex determination in Drosophila melanogaster and Musca domestica converges at the level of the terminal regulator doublesex
Sex-determining cascades are supposed to have evolved in a retrograde manner from bottom to top. Wilkins' 1995 hypothesis finds support from our comparative studies in Drosophila melanogaster and Musca domestica, two dipteran species that separated some 120million years ago. The sex-determining cascades in these flies differ at the level of the primary sex-determining signal and their targets, Sxl in Drosophila and F in Musca. Here we present evidence that they converge at the level of the terminal regulator, doublesex (dsx), which conveys the selected sexual fate to the differentiation genes. The dsx homologue in Musca, Md-dsx, encodes male-specific (MdDSXM) and female-specific (MdDSXF) protein variants which correspond in structure to those in Drosophila. Sex-specific regulation of Md-dsx is controlled by the switch gene F via a splicing mechanism that is similar but in some relevant aspects different from that in Drosophila. MdDSXF expression can activate the vitellogenin genes in Drosophila and Musca males, and MdDSXM expression in Drosophila females can cause male-like pigmentation of posterior tergites, suggesting that these Musca dsx variants are conserved not only in structure but also in function. Furthermore, downregulation of Md-dsx activity in Musca by injecting dsRNA into embryos leads to intersexual differentiation of the gonads. These results strongly support a role of Md-dsx as the final regulatory gene in the sex-determining hierarchy of the housefl
Long-term effectiveness of 6 mm micro-rough implants in various indications: A 4.6- to 18.2-year retrospective study.
OBJECTIVES
To evaluate the long-term effectiveness of 6 mm implants in various indications with a micro-rough surface after 4.6-18.2 years in function and to assess key factors associated with implant survival, success, and biologic/technical complications.
MATERIALS AND METHODS
Fifty-five patients with seventy-four 6 mm implants placed from 2000 to 2013 attended the re-examination assessing well-established clinical and radiographic parameters, biologic and prosthetic complications, and patient-reported outcome measures.
RESULTS
Five implants were lost after a mean follow-up period of 9.1 years resulting in a survival rate of 93.2%. All losses occurred in free-end situations in the mandible. Smoking habit significantly reduced implant survival (hazard ratio 36.25). Two implants exhibited a history of peri-implantitis, and one implant showed progressive marginal bone loss (MBL) resulting in a success rate of 89.2%. The mean MBL amounted to 0.029 mm. Increased MBL was found for implants placed in the maxilla (0.057 mm) and for implants with a diameter of 4.1 mm (0.043 mm). Soft tissue thickness (1.39 mm) and width of keratinized mucosa (1.91 mm) had no effect on MBL. Patient-reported outcome measures showed high satisfaction (mean VAS scores 88%) and high quality of life (mean OHIP-G14 score 2.2).
CONCLUSION
The present study demonstrated survival and success rates of 93.2% and 89.2% for 6 mm implants used in various indications. A factor leading to higher implant failure was smoking, whereas modulating factors increasing annual MBL included implants placed in the maxilla and implants with a diameter of 4.1 mm compared to 4.8 mm
Bone conditioned media (BCM) improves osteoblast adhesion and differentiation on collagen barrier membranes
Background: The use of autogenous bone chips during guided bone regeneration procedures has remained the gold standard for bone grafting due to its excellent combination of osteoconduction, osteoinduction and osteogenesis. Recent protocols established by our group have characterized specific growth factors and cytokines released from autogenous bone that have the potential to be harvested and isolated into bone conditioned media (BCM). Due to the advantageous osteo-promotive properties of BCM, the aims of the present study was to pre-coat collagen barrier membranes with BCM and investigate its effect on osteoblast adhesion, proliferation and differentiation for possible future clinical use.
Methods: Scanning electron microscopy (SEM) was first used to qualitative assess BCM protein accumulation on the surface of collagen membranes. Thereafter, undifferentiated mouse ST2 stromal bone marrow cells were seeded onto BioGide porcine derived collagen barrier membranes (control) or barrier membranes pre-coated with BCM (test group). Control and BCM samples were compared for cell adhesion at 8 h, cell proliferation at 1, 3 and 5 days and real-time PCR at 5 days for osteoblast differentiation markers including Runx2, alkaline phosphatase (ALP), osteocalcin (OCN) and bone sialoprotein (BSP). Mineralization was further assessed with alizarin red staining at 14 days post seeding.
Results: SEM images demonstrated evidence of accumulated proteins found on the surface of collagen membranes following coating with BCM. Analysis of total cell numbers revealed that the additional pre-coating with BCM markedly increased cell attachment over 4 fold when compared to cells seeded on barrier membranes alone. No significant difference could be observed for cell proliferation at all time points. BCM significantly increased mRNA levels of osteoblast differentiation markers including ALP, OCN and BSP at 5 days post seeding. Furthermore, barrier membranes pre-coated with BCM demonstrated a 5-fold increase in alizarin red staining at 14 days.
Conclusion: The results from the present study suggest that the osteoconductive properties of porcine-derived barrier membranes could be further improved by BCM by significantly increasing cell attachment, differentiation and mineralization of osteoblasts in vitro. Future animal testing is required to fully characterize the additional benefits of BCM for guided bone regeneration
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