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Objective@#To evaluate the effect of platelet-rich fibrin extract (PRFe) on the adhesion, proliferation and differentiation of MC3T3-E1 cells cultured on the titanium discs.@*Methods@#Samples were divided into experimental group (P) and control group (D). Group P used the α-minimal essential medium (α-MEM) containing PRFe (0.5%), while group D used only the α-MEM. Cell adhesion and cytoskeleton were observed using scanning electron microscopy (SEM) and laser scanning confocal microscopy (LSCM). Methyl thiazolyl tetrazolium (MTT) assay to detect the number of the osteoblasts at 1, 3, 5, 7 d; the activity of alkaline phosphatase (ALP) to detect the differentiation of osteoblast at 1, 3, 5, 7 d; the level of osteogenetic biomarkers core-binding factorα1 (cbfα1) and osteocalcin (OCN) were quantified by quantitative real-time PCR (qRT-PCR) at 3 and 7 d.@*Results@#SEM and LSCM showed that the adhesion and filaments of group P were higher than those of group D at each time point. MTT assay showed that the absorbance were significantly increased in group P (1 d: 0.299±0.002, 3 d: 0.517±0.004, 5 d: 0.810±0.002, 7 d: 1.203±0.011) compared with group D (1 d: 0.198±0.003, 3 d: 0.399±0.002, 5 d: 0.588±0.002, 7 d: 0.897±0.005) at each time points (P<0.05). Furthermore, the ALP activity of group P (1 d: 0.162±0.004, 3 d: 0.289±0.001, 5 d: 0.491±0.006, 7 d: 0.647±0.005) was significantly higher than that of group D (1 d: 0.121±0.003, 3 d: 0.191± 0.006, 5 d: 0.252±0.004, 7 d: 0.365±0.012), (P<0.05). Moreover, the qRT-PCR showed that the Cbfα1 and OCN gene expression in group P (Cfbα1, 3 d: 1.50±0.04, 7 d: 1.94±0.06; OCN, 3 d: 3.37±0.17, 7 d: 3.92± 0.04) were significantly higher than that in group D(Cfbα1, 3 d: 1, 7 d: 1.18±0.13; OCN, 3 d: 1, 7 d: 2.34± 0.09) (P<0.05).@*Conclusions@#PRFe promoted the adhension, proliferation and differentiation of MC3T3-E1 cells on the titanium discs.
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A patient was treated by multiple dental implants,the implanted 6 was adjucent to impacted 8 .Immediately after implanta-tion,4 month and 3 year after implantation the distace between 8 and 6 implant central line was 4.4,3.2 and 2.5 mm,the angle between 8 long axis and 6 implant central line was 42.3°,45.5°and 50.3°.Then 8 was extracted.
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Objective:To evaluate the effect of concentrate growth factor extract(CGFe)on the proliferation and differentiation of MC3T3-E1 osteoblasts.Methods:MC3T3-1 cells were cultured with and without CGFe respectively.At 1 ,3,5 d of culture the pro-liferation of the cells was detected by MTT assay,the activity of alkaline phosphatase (ALP)of the cells was examined by ALP kit;at 1 4,21 d of culture the calcium nodus formation was observed by Alizarin red staining;RUNX2 and OSX expression was quantified by real-time PCR.Results:With the extension of culture time,the proliferation,ALP activity,the calcium nodus and the level of RUNX2 and OSX mRNA of the experimental group were increased more than those of the control group.Conclusion:CGFe may pro-mote the proliferation and differentiation of osteoblasts.
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<p><b>OBJECTIVE</b>To evaluate the success rate of dental implants and the changes of sinus membrane thickness after sinus lift in cases of thickened sinus membrane.</p><p><b>METHODS</b>Sixteen patients without maxillary sinusitis and with 5-8 mm residual alveolar bone heights were included in this study. The sinus membrane thickeness of these patients were more than 2 mm. All patients received sinus lift surgery and dental implants insertion. The changes of sinus membrane was evaluated by cone-beam CT (CBCT) pre-surgery and 6 months after sinus lift surgery, and the short term success rate of dental implants was also evaluated.</p><p><b>RESULTS</b>A total of 18 implants from 16 subjects were inserted. The thickness of membrane was decreased in 14 cases after sinus lift and increased in 2 cases. All the orifice of maxillary sinus was unobstructed before surgery, one case was obstructed after surgery without inflammation. All the dental implants succeeded.</p><p><b>CONCLUSIONS</b>Sinus augmentation with thickened sinus membrane is not contraindication of sinus lift.</p>
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Humans , Cone-Beam Computed Tomography , Contraindications , Dental Implantation, Endosseous , Dental Implants , Maxilla , Maxillary Sinus , Pathology , General Surgery , Maxillary Sinusitis , General Surgery , Mucous Membrane , Diagnostic Imaging , Pathology , Sinus Floor AugmentationABSTRACT
Objective:To investigate the influence of p38MAPK inhibitor SB203580 on osteoblast MC3T3-E1 cells under high glu-cose concentration.Methods:MC3T3-E1 cells were exposed to glucose at 5.5 mmol/L,25.5 mmol/L and 25.5 mmol/L combined with SB203580 respectively.Cell proliferation was analysed by MTT assay,calcified nodules were quantitated by Alizarin Red S stai-ning.Osteogenesis gene(OCN,Runx2)mRNA level was examined by realtime PCR.Results:High glucose decreased cell prolifera-tion(P<0.05),ALP activity(P<0.05),calcified nodule formation and osteogenesis gene expression of MC3T3-E1 cells.SB203580 reversed the down-regulation of osteogenic markers under high glucose,increased the ALP expression but decreased cell proliferation. Conclusion:SB203580 is antagonistic to P38MAPK in the regulation of preliferation and differetiation of MC3T3-E1 cells.
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<p><b>OBJECTIVE</b>To evaluate the effect of concentrated growth factor extract (CGFe) on the proliferation and differentiation of MC3T3-E1 osteoblasts attached to sandblasted and acid etched titanium surfaces.</p><p><b>METHODS</b>Trials were divided into experimental and control groups. The experimental group used a-MEM that contained CGFe (10% FBS), whereas the control group only used a-MEM (10% FBS). MTT assay was employed to detect the number of osteoblasts on the first, third, and fifth days. Alkaline phosphatase (ALP) activity and scanning electron microscope (SEM) were used to detect the osteoblast differentiations on the third and fifth days and to observe the osteoblast extensions on titanium surfaces for 12 h, respectively. Meanwhile, the levels of the osteogenetic biomarkers Runt-related transcription factor-2 (Runx2) and Osterix (Osx) on the third and seventh days were quantified via real-time polymerase chain reaction (PCR).</p><p><b>RESULTS</b>MTT assay indicated that on the first, third, and fifth days, the absorbance in the experimental group significantly increased than that in the control group (P < 0.05). ALP activity: on the third and fifth days, the absorbance of the experimental group was significantly higher than that of the control group (P < 0.05). SEM: at 12 h, the extension of the experimental group cells was larger than that of the control group. Real-time PCR: given the standardization in the group, the gene expression level of the control group on the third day was 1, and the Runx2 and Osx gene expressions in the experimental group were larger than those of the con- trol group.</p><p><b>CONCLUSION</b>CGFe can efficiently stimulate the proliferation, differentiation and extension of MC3T3-E1 cells.</p>
Subject(s)
Cell Differentiation , Cell Line , Cell Proliferation , Core Binding Factor Alpha 1 Subunit , Intercellular Signaling Peptides and Proteins , Osteoblasts , Osteogenesis , TitaniumABSTRACT
Objective:To study the osseointegration of implant in fresh extraction socket with or without bone grafting.Methods:In the mandibular premolar region of 6 beagle dogs,bone defect in the size of (3~4)mm ×(3~4)mm ×(5 ~6)mm on the mesial wall of the mesial root socket was made.On control side implants were installed immediately into the extraction sockets(group A).On an-other side Bio-Oss grafts and membrane(GBR)were placed following implantation(group B).All animals were sacrificed 3 months af-ter implantation,specimens were examined for histo-morphometric analysis of bone to implant contact and new bone formation.Results:No implant was loosening in the 2 groups.New bone was filled in the bone defect areas in 2 groups.No statistical difference of the per-centage of new bone formation and bone-to-implant contact ration(BIC)was observed between 2 groups.Conclusion:With the defect in a certain size on the root socket wall osseointegration may occur between the new bone and implant without bone transplantation.
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Objective To test the safety and effectiveness of transpedicular fixation combined with transpedicular bone grafting via less invasive paraspinal intermusclar approach in treatment of thoracolumbar fractures.Methods The study involved 23 cases of thoracolumbar fractures treated with paraspinal multifidus intramusclular mini-incision,transpedicular bone grafting,and short-segment pedicle screw fixation from June 2009 to June 2012.There were 16 males and 7 females at age of 19-55 years (average 38.8years).Time from injury to surgery varied from 6 hours to 7 days (average 3.2 days).Fracture level was T11 in three cases,T12 in seven,L1 in nine,and L2 in four.According to Denis fracture classification,there were altogether 10 compression fractures and 13 burst fractures.McCormack load sharing classification scored average 5.3.Before operation,anterior vertebral body height ratio was average 58.6％(range,45 ％-73％) and kyphosis angle was average 23.7° (range,15°-34°).Results Operation lasted for average 95.5 minutes (range,75-130 minutes) with intraoperative bleeding of average 160.3 ml (range,115-220 ml).Unilateral incision that was averaged 3.5 cm (range 3.2-4.0 cm) in length obtained primary healing.Average follow-up time was 12.6 months (range,7.5-18 months).Average height of the anterior border was corrected to 97.3％ and average kyphosis angle was corrected to 4.6°.There was neither instrumentation failure nor symptom of persistent postoperative back pain.Conclusions Transpedicular fixation with transpedicular bone grafting via paraspinal muscle approach provides effective recovery of vertebral morphology and correction of kyphotic deformity.Furthermore,the technique gains advantages of easy operation,small trauma,less blood loss and rapid recovery.
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<p><b>OBJECTIVE</b>To study mRNA expression of receptor activator nuclear factor kappa B ligand (RANKL) and its decoy receptor, osteoprotegerin (OPG) in peri-implant tissue during unloading period.</p><p><b>METHODS</b>An animal model of dental implant was established in 6 male Beagle dogs of 1-2 years old. Bone remodeling was tested at 3, 7, 15, 30, 60 and 90 days since the placement of implants. RANKL and OPG mRNA expression were quantified by real-time polymerase chain reaction (PCR). Then mandibular bones were taken out and the morphological changes were observed by X-ray, bone tissue was tested by immunohistochemistry stain.</p><p><b>RESULTS</b>The most prominent period of bone remodeling occurred at 7th day after the placement of implants. The expression of RANKL and OPG increased in a time-dependent manner in both soft and hard tissue. After 7 days they gradually decreased.</p><p><b>CONCLUSION</b>RANKL and OPG can express in soft tissue, and the changing tendency is consistent with the change of bone remodeling, it indicates that RANKL and OPG play an important role in the bone remodeling.</p>
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Animals , Dogs , Male , Bone Remodeling , Bone and Bones , Carrier Proteins , NF-kappa B , Osteoprotegerin , RANK Ligand , Receptor Activator of Nuclear Factor-kappa BABSTRACT
Objective To observe the effects of transplantation of bone marrow stem cells(BMSCs)transfected with bone morphogenetic protein-2(BMP-2)on fracture healing in rats with diabetes so as to provide a new therapy for diabetic fractures.Methods Fifty male adult Wistar rats aged six weeks randomized to the control group and experimental group were all employed to establish models with diabetic fractures.Under high glucose condition,BMSCs were transfected with BMP-2 by adenovirus vector in vitro.BMSCs transfected by BMP-2 were transplanted into the fracture area of rats in the experimental group,while non-transfected BMSCs into the corresponding area of rats in the control group.X-ray examination was performed at 1,2,3,4 and 6 weeks after transplantation.Bony calluses were collected for HE staining and gray scales of BMP-2 in calluses were determined by immunohistochemical method.Meanwhile,serum levels of BMP-2 were measured by ELtSA.Results The gray scales of BMP-2 in the calluses were 83±3 in the experimental group and 118±4 in the control group at four weeks(P＜0.01).The serum concentrations of BMP-2 were(203.80±8.96)ng/L in the experimental group and(139.15±4.19)ng/L in the control group at four weeks(P＜0.01).Conclusion Transplantation of BMSCs transfected by BMP-2 promotes fracture healing in diabetic rats.
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Objective To observe the effects of olfactory lamina propria (OLP) transplantation and ganglioside GM1 treatment on spinal cord injury (SCI) in rats.Methods Totally 50 healthy pure breed female Sprague-Dawley (SD) rats after spinal cord hemiseetion were randomly divided into 5 groups and were given different treatments: (OLP + GM1) treatment group (group A), GM1 treatment group (group B), OLP treatment group (group C), spinal cord injury but without treatment group (group D) and healthy control group (group E). The recovery of neurological function was evaluated by somatosensory evoked potential (SEP) and pathological examination after surgery. Results In group A, in some rats an escaping response in right hind leg occurred, but in other groups, the motor function was not significantly improved. Histological examination showed that transplanted olfactory lamina propria survived in the transplantation area and expanded on certain routes. NF positive nerve fibers passed through the transplantation area. Compared with group B, C, D, the N1-wave latency was(4.71±0. 72)ms 4 weeks after operation(P<0. 01), and the NF density was(7. 31±0. 26) ×104/mm28 weeks after operation in group A(P<0. 05). Conclusions Olfactory lamina propria (OLP) transplantation and ganglioside GM1 treatment have a synergistic effect on SCI.
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Twelve Sprague-Dawley albino rats (24 eyes) were used in this study. Nor- mal saline was injected into the anterior chamber of the experimental eye until the b-wave of the electoretinogram was extinguished, then maintained the critical high intraocular pressure (HIOP) for 90 minutes. Just a puncture, without HIOP, was given to the anterior chamber of the other eye of the same rat as a control Both retinas of the experimental and control eyes were whole-mounted and treated with succinate dehydrogenase (SDH) histochemical technique. The reacted products of SDH showed in either retinal ganglion cells (RGCs) or optic nerve fibers of the experimental eyes were lighter than those of the control eyes under microscope. Furthermore, the grey scale values of the reacted products of SDH on the matched ratinas were measured at corresponding microscopic fields and RGCs with an image analysis system. The differences of the grey scale va- lues between the experimental and control retinas were high statistically signi- ficant (p
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Rapid Golgi preparations of golden hamster dorsal lateral geniculate nucleus (LGd)show three classes of neurons with different morphological characteristics. Class Ⅰ and class Ⅲ cells are distributed throughout the LGd.Class Ⅱ cells are found more common medially at mid-genieulate levels.Class Ⅰ cells are multipolar neurons with large cell body.The smooth primary dendrites branch extensively. Spine-like and small excrescences are found along distal dendrites.Class Ⅱ cells have medium size pear-like cell body.The distal dendrites are usually derived from one primary dentrite.Dense cluster of dentritic protrusions is usually found at branch points of the dendrites.Class Ⅲ cells have the smallest cell body and are paucidendritie but the dendrites have complex protrusions including pearl-like dendritic appendages.Class Ⅰ and class Ⅱ cells possess an axon but most of class Ⅲ cells are not found to have an axon. The connections between the LGd and visual cortex was investigated using the retrograde horseradish peroxidase(HRP)histochemical method.After injection of HRP into the visual cortex,Class Ⅰ and class Ⅱ neurons were labelled but class Ⅲ celts were not suggesting that class Ⅰ and class Ⅱ cells are projecting neurons.Class Ⅲ cells do not seem to project to the visual cortex and are probably local circuit neurons.
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The changes of cytochrome oxidase activity of retinal ganglion cell (RGC) caused by the artificial high intraocular pressure (HIOP) was presented in this paper. Normal saline was injected into the anterior chamber of the right eye of 36 rats. that resulted in an intraocular pressure up to 60 mmHg for 3 hours. The experimental rats were divided into 0-, 3- and 7-day groups according to their survival time. Both retinae of each rat were whole mounted on the same slide, the left one being used as control. Thirty-six pairs of retina were treated with the modified cytochrome oxidase (CCO) histochemical technique under similar conditions. The high CCO activity of RGC were counted. The extinction of the cytoplasm of RGC, which indicates the degree of CCO activity, was measured with microphotometer. As compared with the normal eyes, the high CCO activity RGC of experimental eyes were reduced significantly. It was found that the high CCO activity of RGC in the temporal side of retina has been reduced much more than that of the nasal side. However, the high CCO activity of RGC in 3- and 7-day groups were more than that of 0-day group, the extinction of CCO activity of RGC in 7-day group was lower than that of the 3-day group. These facts showed that the CCO activity might restore in various degrees followed by a longer survival time. This experiment emphasized the fact that the HIOP led to metabolic changes of RGC, which may be of value to the study of glaucoma.