OBJECTIVE: To explore the difference of bone formation potential between hypertrophic nonunion tissue and atrophic nonunion tissue, which may be beneficial to nonunion therapy. METHODS: From October 2010 to March 2014, 40 nonunion tissue samples were collected in Department of Orthopedics, Second Xiangya Hospital. The samples were divided into a hypertrophic nonunion group (n=20) and an atrophic nonunion group (n=20) according to nonunion character; or a 20 to 35 years old group (n=18), a 36 to 50 years old group (n=18), a more than 50 years old group (n=4) according to different ages; or a 9-12 months group (n=21), 13-24 months group (n=14) and a more than 24 months group (n=5) according to different nonunion time. Semi-quantification was performed by SP immunohistochemical method and IPP6.0 was used to analyze the expression of bone morphogenetic protein-2 (BMP-2) through measuring the mean optical density. RESULTS: The mean optical density of BMP-2 was 0.1540±0.0408 in hypertrophic nonunion tissue, 0.1372±0.0372 in atrophic nonunion tissue, there was no significant difference between the 2 groups (P>0.05). The mean optical density of BMP-2 was 0.1477±0.0379 in the 20 to 35 years old group, 0.1419±0.0399 in the 35 to 50 years old group, 0.1456±0.0595 in the more than 50 years old group, there was no significant difference among the three groups (P>0.05). The mean optical density of BMP-2 was 0.1449±0.0366 in the 9-12 months group, 0.1472±0.0400 in the 13-24 months group, 0.1445±0.0541 in the more than 24 months group, there was no significant difference among the 3 groups (P>0.05). CONCLUSION The present results suggest that the hypertrophic nonunion tissue share similar osteogenic potential with the atrophic nonunion tissue.
Adult ; Bone Morphogenetic Protein 2 ; genetics ; metabolism ; Fracture Healing ; Fractures, Ununited ; genetics ; metabolism ; Humans ; Middle Aged ; Osteogenesis ; Young Adult

Pentaxin 3 (PTX3), as a multifunctional glycoprotein, plays an important role in regulating inflammatory response, promoting tissue repair, inducing ectopic calcification and maintaining bone homeostasis. The effect of PTX3 on bone mineral density (BMD) may be affected by many factors. In PTX3 knockout mice and osteoporosis (OP) patients, the deletion of PTX3 will lead to decrease of BMD. In Korean community "Dong-gu study", it was found that plasma PTX3 was negatively correlated with BMD of femoral neck in male elderly patients. In terms of bone related cells, PTX3 plays an important role in maintaining the phenotype and function of osteoblasts (OB) in OP state;for osteoclast (OC), PTX3 in inflammatory state could stimulate nuclear factor κ receptor activator of nuclear factor-κB ligand (RANKL) production and its combination with TNF-stimulated gene 6(TSG-6) could improve activity of osteoclasts and promote bone resorption;for mesenchymal stem cells (MSCs), PTX3 could promote osteogenic differentiation of MSCs through PI3K/Akt signaling pathway. In recent years, the role of PTX3 as a new bone metabolism regulator in OP and fracture healing has been gradually concerned by scholars. In OP patients, PTX3 regulates bone mass mainly by promoting bone regeneration. In the process of fracture healing, PTX3 promotes fracture healing by coordinating bone regeneration and bone resorption to maintain bone homeostasis. In view of the above biological characteristics, PTX3 is expected to become a new target for the diagnosis and treatment of OP and other age-related bone diseases and fracture healing.
Animals ; Male ; Mice ; Bone Resorption/metabolism* ; Cell Differentiation ; Fracture Healing/genetics* ; Osteoblasts ; Osteoclasts ; Osteogenesis ; Osteoporosis/genetics* ; Phosphatidylinositol 3-Kinases/pharmacology*

OBJECTIVETo investigate the expression change of integrin alpha 5 during mandibular fracture healing.

METHODSUsing rabbit mandibular fracture model, the fractured bone tissues were obtained and paraffin slices were made on the days of 1, 3, 5, 7, 14, 30, 60 and 90 after surgery, respectively. Non-fractured mandibles were used as normal control. LsAB immunohistochemical method was used to detect the expression of integrin alpha 5 in bone tissue, especially on osteoblasts and osteoclasts.

RESULTSIntegrin alpha 5 was widely expressed in fractured bone and surrounding soft tissues. The expression of integrin alpha 5 increased 7 days after fracture, peaked in 14 to 30 days and returned to nearly normal in 60-90 days.

CONCLUSIONDuring mandibular fracture healing, the expression of integrin alpha 5 in bone tissue increased clearly. It is estimated that integrin alpha 5 players an important role in fracture healing.

Animals ; Female ; Fracture Healing ; Immunohistochemistry ; In Situ Hybridization ; Integrin beta1 ; biosynthesis ; genetics ; Male ; Mandibular Fractures ; metabolism ; Osteoblasts ; physiology ; RNA, Messenger ; biosynthesis ; genetics ; Rabbits

OBJECTIVETo explore the effect of fluvastatin on vascular endothelial growth factor (VEGF) in rats with osteoporosis in the process of fracture healing.

METHODSFractures at the intermediate piece of the femur were made on 72 Sprague Dawley (SD) rats (weighing initially 290-340 g and aged 6 months) with osteoporosis after ovariectomy for three months, then these rats were divided randomly into the medication administration group (the experimental group) and the control group, 36 rats each. In the experimental group, the rats received fluvastatin lavage (10 mg/kg per day) since the next day of operation lasting for 6 weeks, and the rats in the control group received placebo. Then the expression of VEGF and VEGF mRNA in bony callus of the two groups was measured respectively with immunohistochemistry and in situ hybridization on days of 3rd, 7th, 14th, 21st, 28th, and 42nd, and image analysis was made with real-color image analysis machine.

RESULTSNo difference was found in the cellular localization of VEGF and VEGF mRNA gene expression between the experimental group and the control group in process of fracture healing and their expression modes were almost similar. On the 14th day postoperatively, the positive extent of positive cells in the experimental group was higher than that of the control group (P < 0.05).

CONCLUSIONFluvastatin can promote the VEGF level in rats with osteoporosis in process of fracture healing.

Animals ; Fatty Acids, Monounsaturated ; pharmacology ; Fracture Healing ; Immunohistochemistry ; In Situ Hybridization ; Indoles ; pharmacology ; Osteoporosis ; metabolism ; RNA, Messenger ; analysis ; Rats ; Rats, Sprague-Dawley ; Vascular Endothelial Growth Factor A ; analysis ; genetics

OBJECTIVE: To explore effect of tobramycin (TOB) on healing of femoral fractures in rats. METHODS: Totally 32 male sprague-dawley (SD) rats were selected and randomly divided into sham group (group A), fracture group (group B), fracture with TOB group (group C) and fracture + TOB + IWR-1 group (group D), 8 rats in each group. Close femoral fracture model in rats were established in group B, C and D, group A was sham operation without otherwise process. Group D was intraperitoneal injected 100 μl (8 μM) of Wnt pathway inhibitor IWR-1-endo (IWR-1) before molding at 1 day. At 1 day after molding, 100 μl (100 μM) of TOB was intraperitoneally injected into group C and D at once a day for 7 days. At 7 weeks after modling, fracture healing of group B, C and D were observed by X-ray, Western blotting was appilied to detect alkaline phosphatase(ALP) and Runt related transcription factor 2 (RUNX2) and β-catenin of Wnt passway. RESULTS: X-ray results showed fracture line disappeared, callus formation and fracture healing well in group C compared with begning of molding; while a little fracture line, callus formation and fracture malunion in group B and d could be seen. Western blotting results showed ALP, RUNX2 and expression of β-catenin in group B, C and D were higher than that of group A ( CONCLUSION Tobramycin could promote osteoblast differentiation and fracture healing by stimulating Wnt / β-catenin signaling pathway, up regulating expression of ALP and RUNX2.
Alkaline Phosphatase ; Animals ; Cell Differentiation ; Core Binding Factor Alpha 1 Subunit/genetics* ; Femoral Fractures ; Fracture Healing ; Male ; Osteogenesis ; Rats ; Tobramycin ; Wnt Signaling Pathway ; beta Catenin/metabolism*

To compare the effects between gene therapy and gradual release carrier for bone morphogenetic protein-2 (BMP-2) in repairing bone defects, bone defects for 15 mm were created.on the bilateral radius in rabbits and treated with four kinds of implantations, ie, composite of transgeneic MSCs and PLA/PCL (Group A), composite of MSCs and gradual release carrier for BMP-2 (Group B), composite of MSCs and PLA/PCL (Group C), and PLA/PCL alone (Group D). After 4, 8, and 12 weeks of the operations, X-ray, histological examination, biomechanics analysis, and bone density measurement were conducted. Results showed that both osteoblasts and mesenchymal cells displayed strongly positive expression of BMP-2 in Group A after 4 weeks of the operation, the speed and quality of bone formation in Group A were much better than those in Group B. After 12 weeks of the operations, bone defects were completely repaired in Group A. BMP-2 gene therapy is really a good method to repair segmental bone defects.
Animals ; Bone Morphogenetic Protein 2 ; genetics ; therapeutic use ; Caprolactam ; analogs & derivatives ; chemistry ; Fracture Healing ; Genetic Therapy ; Implants, Experimental ; Lactic Acid ; chemistry ; Mesenchymal Stem Cell Transplantation ; methods ; Osteogenesis ; Polyesters ; Polymers ; chemistry ; Rabbits ; Radius Fractures ; therapy ; Tissue Scaffolds

OBJECTIVETo study the effect of poly D,L-lactic acid (PDLLA) screws with PDLLA-rhBMP compound on bone regeneration in the screw holes and fracture ends of dog mandibles.

METHODSA self-control study was carried out in 4 dogs. PDLLA/rhBMP-2 compound screws were implanted to fix the mental fractures and PDLLA screws were used as control. The samples from mandibles were collected at 4, 8, 12, 16 weeks after implantation and observed by radiography and histology.

RESULTSAll dogs showed a greater degree of bone regeneration around PDLLA/rhBMP-2 screws than PDLLA ones and all fractures were fixed and healed well.

CONCLUSIONThe PDLLA-rhBMP screw has a better effect of inducing osteogenesis than PDLLA screw, and is able to exert a good fixation to fracture.

Animals ; Biocompatible Materials ; administration & dosage ; Bone Morphogenetic Protein 2 ; Bone Morphogenetic Proteins ; administration & dosage ; biosynthesis ; genetics ; Bone Screws ; Dogs ; Fracture Fixation, Internal ; methods ; Fracture Healing ; Lactic Acid ; administration & dosage ; Male ; Mandibular Fractures ; surgery ; Polyesters ; Polymers ; administration & dosage ; Recombinant Proteins ; administration & dosage ; biosynthesis ; genetics ; Transforming Growth Factor beta

Adenovirus carrying BMP-2 gene, after being mixed with fibrinous gel, was siphoned off on biodegradable scaffolds (PLA/PCL). The composite was used to repair 1.5 cm long radius defect in rabbits. Four methods were in use in the experiments: Ad-BMP-2 plus fibrinous gel and PLA/PCL (Group A), reconstructed hBMP-2 plus fibrinous gel and PLA/PCL (Group B), Ad-Lacz plus fibrinous gel and PLA/PCL (Group C), and fibrinous gel and PLA/PCL (Group D). Results showed that the defects treated in Group A were repaired with much more new bone regenerated, bridged earlier and stronger than those in Group B 12 weeks after operation. The defects treated in the other two groups could not attain osseous tissue healing. BMP-2 gene carried by biodegradable scaffold and fibrinous gel is easy to conduct and has very strong osteoinduction ability. It is really a good method to repair segmental bone defects.
Absorbable Implants ; Adenoviridae ; genetics ; metabolism ; Animals ; Biocompatible Materials ; Bone Morphogenetic Protein 2 ; biosynthesis ; genetics ; Bone Substitutes ; Caprolactam ; analogs & derivatives ; chemistry ; Fracture Healing ; Gels ; Genetic Therapy ; Humans ; Implants, Experimental ; Lactic Acid ; chemistry ; Polyesters ; Polymers ; chemistry ; Rabbits ; Radius Fractures ; therapy ; Tissue Engineering ; Tissue Scaffolds