Necrosulfonamide reverses pyroptosis-induced inhibition of proliferation and differentiation of osteoblasts through the NLRP3/caspase-1/GSDMD pathway.

The acute inflammatory stimulation occurring after a bone fracture regulates the repair and healing of local bone injury; however, under certain conditions, pyroptosis may occur in osteoblasts, which affects osteoblast proliferation and differentiation, thereby affecting the growth, development and morphological changes of bone tissue. The aim of the present study was to examine the effect of the pyroptosis inhibitor necrosulfonamide (NSA) on the proliferation and differentiation of osteoblasts and elucidate the underlying mechanism. The results revealed that NSA reversed the effects of ATP/lipopolysaccharide (LPS) on cell viability and pyroptosis, and on the mRNA and protein expression of pyroptosis-related genes. It also suppressed the secretion of IL-6, TNF-α and IL-1ß and reversed the effects of ATP/LPS on the activity of ALP and the mRNA expression of differentiation-related genes in osteoblasts. The fact that overexpression of caspase-1, gasdermin D (GSDMD) and NLRP3 abolished the effects of NSA on the viability and pyroptosis of osteoblasts, as well as the mRNA expression of differentiation-related genes and the activity of ALP in osteoblasts, indicated that NSA promoted the proliferation and differentiation of osteoblasts by inhibiting the NLRP3/caspase-1/GSDMD pyroptosis pathway. The present study provides proof supporting the potential application of NSA for improving the function of osteoblasts in fracture repair and indicates the value of the NLRP3/caspase-1/GSDMD pyroptosis pathway as a pharmaceutical target.

Actin polymerization state regulates osteogenic differentiation in human adipose-derived stem cells.

BACKGROUND: Actin is an essential cellular protein that assembles into microfilaments and regulates numerous processes such as cell migration, maintenance of cell shape, and material transport. METHODS: In this study, we explored the effect of actin polymerization state on the osteogenic differentiation of human adipose-derived stem cells (hASCs). The hASCs were treated for 7 days with different concentrations (0, 1, 5, 10, 20, and 50 nM) of jasplakinolide (JAS), a reagent that directly polymerizes F-actin. The effects of the actin polymerization state on cell proliferation, apoptosis, migration, and the maturity of focal adhesion-related proteins were assessed. In addition, western blotting and alizarin red staining assays were performed to assess osteogenic differentiation. RESULTS: Cell proliferation and migration in the JAS (0, 1, 5, 10, and 20 nM) groups were higher than in the control group and the JAS (50 nM) group. The FAK, vinculin, paxillin, and talin protein expression levels were highest in the JAS (20 nM) group, while zyxin expression was highest in the JAS (50 nM) group. Western blotting showed that osteogenic differentiation in the JAS (0, 1, 5, 10, 20, and 50 nM) group was enhanced compared with that in the control group, and was strongest in the JAS (50 nM) group. CONCLUSIONS: In summary, our data suggest that the actin polymerization state may promote the osteogenic differentiation of hASCs by regulating the protein expression of focal adhesion-associated proteins in a concentration-dependent manner. Our findings provide valuable information for exploring the mechanism of osteogenic differentiation in hASCs.

Anatomic measurement of osseous parameters of the glenoid.

The angle and position of the scapular glenoid are important in shoulder mechanics, the interpretation of diseases, and planning shoulder replacement surgery. In total shoulder replacement, understanding the bony parameters of the glenoid is also of considerable guiding significance for designing implant size and improving material adaptability. To compare glenoid parameters measured from skeletal scapula specimens with those measured by 3D modeling of CT scanning images, analyze correlations between these data, and draw conclusions to guide clinical treatment of shoulder joint injury and total shoulder joint replacement. The data of manual and CT measurements from the same Chinese dry glenoid was compared. Three-dimensional measurement data were collected from the Japanese population and compared with the Chinese population data generated in this study. There were no significant differences between manual measurement and CT measurement in the inclination angle, glenopolar angle, anteroposterior transverse diameter, upper to lower vertical diameter, and depth of the glenoid (P = 0.288, 0.524, 0.111, 0.194, and 0.055, respectively). Further, there were no significant differences between Japanese and Chinese glenoid bones in the upper and lower vertical diameters or anteroposterior transverse diameters (P > 0.05). There were no significant differences between CT and manual measurements, suggesting that the CT method may provide measurements very close to the actual specimen size. This result, however, indicated that the measurer should be careful when measuring the depth of the glenoid.

[Application of picture archiving and communication systems in lateral wedge osteotomy for cubitus varus deformity in teenagers].

OBJECTIVE: To investigate effectiveness of picture archiving and communication systems (PACS) in lateral wedge osteotomy for cubitus varus deformity in teenagers. METHODS: A clinical data of 16 teenagers with cubitus varus deformity between July 2014 and July 2016 was retrospectively analyzed. All patients were treated with lateral wedge osteotomy and fixed with plate. Before operation, the osteotomy design (the osteotomy angle and length) was done in the PACS, including the carrying angle of healthy limb and the varus angle of affected side. There were 10 males and 6 females, with an average age of 11.4 years (range, 10-17 years). The disease duration ranged from 2 to 10 years (mean, 5.6 years). The preoperative X-ray film showed that the supracondylar fractures of the humerus had all healed, and 9 cases had internal rotation deformity; the varus angle of the affected side was 19.5°-33.5°. After operation, the fracture healing and cubitus varus deformity correction were observed by X-ray films, the elbow function was evaluated by Mayo scoring, and the elbow range of motion was detected. RESULTS: There was no significant difference between the actual intraoperative osteotomy angle and length and the preoperative design ( P>0.05). The hospital stay was 2-8 days, with an average of 4.5 days. No complication such as incision infection or ulnar nerve injury occurred. All 16 cases were followed up 12-18 months, with an average of 14 months. X-ray films showed that the osteotomy healed at 2-7 months after operation, with an average of 2.5 months. The internal fixators were removed within 8-14 months after operation (mean, 12.0 months). X-ray films measurement showed that the carrying angle of the affected side recovered to (10.3±2.0)° at 1 day after operation, which was not significantly different from that of the healthy side [(10.6±1.5)°] before operation ( t=0.480, P=0.637). The carrying angle of the affected side was (9.8±2.6)° at 1 year after operation, which was not significantly different from that of the healthy side [(10.4±1.6)°] at the same time point ( t=0.789, P=0.438). At 1 year after operation, the ranges of flexion and extension of affected side were (131.6±8.4)° and (6.4±2.6)°, respectively; and the ranges of flexion and extension of healthy side were (134.2±6.3)° and (5.9±2.2)°, respectively. There was no significant difference between the healthy and affected sides ( t=1.143, P=0.262; t=0.587, P=0.561). The elbow joint function at 1 year after operation evaluated by Mayo scoring standard rated as excellent in 9 cases, good in 6 cases, and fair in 1 case, and the excellent and good rate was 93.7%. CONCLUSION: Before lateral wedge osteotomy, the PACS is used to design the osteotomy angle and length, which can guide the operation and make the osteotomy more accurate and simple.

Mechanical Sensing Element PDLIM5 Promotes Osteogenesis of Human Fibroblasts by Affecting the Activity of Microfilaments.

Human skin fibroblasts (HSFs) approximate the multidirectional differentiation potential of mesenchymal stem cells, so they are often used in differentiation, cell cultures, and injury repair. They are an important seed source in the field of bone tissue engineering. However, there are a few studies describing the mechanism of osteogenic differentiation of HSFs. Here, osteogenic induction medium was used to induce fibroblasts to differentiate into osteoblasts, and the role of the mechanical sensitive element PDLIM5 in microfilament-mediated osteogenic differentiation of human fibroblasts was evaluated. The depolymerization of microfilaments inhibited the expression of osteogenesis-related proteins and alkaline phosphatase activity of HSFs, while the polymerization of microfilaments enhanced the osteogenic differentiation of HSFs. The evaluation of potential protein molecules affecting changes in microfilaments showed that during the osteogenic differentiation of HSFs, the expression of PDLIM5 increased with increasing induction time, and decreased under the state of microfilament depolymerization. Lentivirus-mediated PDLIM5 knockdown by shRNA weakened the osteogenic differentiation ability of HSFs and inhibited the expression and morphological changes of microfilament protein. The inhibitory effect of knocking down PDLIM5 on HSF osteogenic differentiation was reversed by a microfilament stabilizer. Taken together, these data suggest that PDLIM5 can mediate the osteogenic differentiation of fibroblasts by affecting the formation and polymerization of microfilaments.

Distribution and property of nerve fibers in human long bone tissue.

OBJECTIVE: To observe the distribution of the nerve fibers in the bone tissue and the entry points of these fibers into the bone. METHODS: The adult tibia was used for the ground sections which were afterwards made into the slice sections by decalcification in ethylenediamine tetraacetic acid (EDTA). The ground sections were stained in silver and the slice sections were stained in silver and haematoxylin and eosin (HE) respectively. Then, the samples of the transmission electron microscope and the atomic force microscope were made and observed. RESULTS: In the human long bone tissue, many nerve fibers were distributed in the membrane, cortical bone, cancellous bone and marrow. The nerve fibers entered the bone from the nutrient foramen, and passed through the nutrient canal, Haversian's canal and Volkmann's canal, and finally into the bone marrow. In the nutrient canal, the nerve fibers, mainly the medullary nerve fibers, followed the blood vessel into the bone. In the cortical bone, the nerve fibers also followed the blood vessels and were mainly distributed along Haversian's canal and Volkmann's canal. In the bone trabecular and bone marrow, there were many nerve fiber endings arranged around the blood vessels, mainly around the tunica media of medium-size arteries in the marrow and around capillary blood vessels, and a few scattered in the bone marrow. There were sporadic nerve endings in epiphyseal plate and no nerve fibers permeated epiphysis to diaphysis. No distribution of nerve fibers could be found in cartilaginous part. CONCLUSIONS: There are many nerve fibers in bone and the nerve passageway is nutrient foramen, Volkman's canal, Haversian's canal and bone marrow.

[In vitro biocompatibility of novel absorbable hydroxyapatite and AO artificial bone beta-tricalcium phosphate with rhesus bone marrow stromal cells].

OBJECTIVE: To study the in vitro biocompatibility of novel hydroxyapatite (HA) and AO artificial bone beta-tricalcium phosphate (beta-TCP) with rhesus bone marrow stromal cells (rBMSCs) . METHODS: The third passage of rBMSCs were cultured with HA and beta-TCP respectively, with the cells cultured without the materials as the control. The morphology and proliferation of cells were observed by inverted phase-contrast microscope and scanning electron microscope (SEM). MTT assay was used to semiquantitatively evaluate the cell proliferation. RESULTS: The rBMSC cocultured with HA exhibited good growth as observed under inverted phase-contrast microscope, without significant difference from the cells in the control group. Some small particles were seen pealing off from beta-TCP, and some of the cells died. Under SEM, rBMSCs showed good adhesion to HA with obvious proliferation, but the ratio of adhesive cells was not as high as that in beta-TCP group. MTT assay showed no significant difference in the cell number between HA and the control groups, but the cell number in beta-TCP group was notably less than that of control group. CONCLUSION: Novel HA has good biocompatibility with rBMSCs for bone tissue engineering, and AO artificial bone still needs improvement to serve as scaffold material for BMSCs.

[Bone defect repair with a new tissue-engineered bone carrying bone morphogenetic protein in rabbits].

OBJECTIVE: To construct a new tissue-engineered bone with poly (D, L-lactide-co-glycolide) (PLGA), bone morphogenetic protein (BMP) and bone marrow-derived stem cells (BMSCs) and observe its effect in repairing segmental bone defects. METHODS: A 15-mm bone defect in the right radius was induced in New Zealand white rabbits, and the models were randomized into three groups to receive implantation of the tissue-engineered bone grafts constructed with PLGA carrying 5 mg BMP and about 1 x 10(6) BMSCs (experimental group), grafts of PLGA with about 1 x 10(6) BMSCs (control group), or grafts of exclusive PLGA (blank control group), respectively. The osteogenesis in the bone defect after the implantation on was evaluated X-ray films, and the histological changes of the tissues sampled from the bone defect 4, 8, and 12 weeks after operation were observed and new bone formation was measured by image analysis. RESULTS: The bone defect was completely repaired in the experimental group 12 weeks after the implantation, showing the best results among the 3 groups. The bone defects in the blank control group was filled with only fibrous and connective tissues at 12 weeks. CONCLUSION: This tissue-engineered bone constructed with PLGA, BMP and BMSCs possesses good ability in repairing segmental bone defect.

Prognostic Value of Ezrin in Various Cancers: A Systematic Review and Updated Meta-analysis.

More and more studies have investigated the effects of Ezrin expression level on the prognostic role in various tumors. However, the results remain controversial rather than conclusive. Here, we performed a systematic review and meta-analysis to evaluate the correlation of Ezrin expression with the prognosis in various tumors. the pooled hazard ratios (HR) with the corresponding 95% confidence intervals (95% CI) were calculated to evaluate the degree of the association. The overall results of fifty-five studies with 6675 patients showed that elevated Ezrin expression was associated with a worse prognosis in patients with cancers, with the pooled HRs of 1.86 (95% CI: 1.51-2.31, P < 0.001) for over survival (OS), 2.55 (95% CI: 2.14-3.05, P < 0.001) for disease-specific survival (DFS) and 2.02 (95% CI: 1.13-3.63, P = 0.018) for disease-specific survival (DSS)/metastasis-free survival (MFS) by the random, fixed and random effect model respectively. Similar results were also observed in the stratified analyses by tumor types, ethnicity background and sample source. This meta-analysis suggests that Ezrin may be a potential prognostic marker in cancer patients. High Ezrin is associated with a poor prognosis in a variety of solid tumors.

A new approach to tissue repair: gene therapy.

The process of tissue repair involves a complex tissue response to injury in which growth factors, playing a major role in this process, trigger, control and terminate soakage of inflammatory cells, cells proliferation, secretion of matrix and scars formation by autocrine, paracrine or both. Thus, growth factors can be used to alter the microenvironment of the wounded tissues and to promote their repair. But, there are notable disadvantages in using purified recombination growth factors, 1) the source is so limited that their prices are expensive; 2) the ir half-lives are short and easy to be destroyed by wound proteases; 3) there is no perfect carrier; 4) high initial doses are required but easy to bring toxicity; 5) it is difficult to apply growth factors in deep wounded tissues again and again, their function cannot be played enough accordingly; 6) most of growth factors are the products of recombination. All above-mentioned disadvantages result in a low activity.

[Dynamic changes of SOD and MDA in canine limbs with gunshot wound in hot and humid environment: an experimental study].

OBJECTIVE: To study the dynamic changes in lipid peroxidation and the activity of antioxidative enzyme in canine limbs with gunshot wound in hot and humid environment. METHODS: Eighteen dogs with gunshot wound were randomly assigned into 3 groups with equal numbers. Dogs observed after injury in normal environment was designated as NE group, those in hot and humid environment as HHE group and those in hot and humid environment with preceding heat acclimatization training as HA group. Contents of superoxide dismutase (SOD) and malonaldehyde (MDA) in both the peripheral blood and muscular tissues from gunshot wound tract were measured at 0, 1, 3, 4, 6, 8, 10, 14, 18 and 24 h respectively after the dogs had been wounded. RESULTS: Positive correlation between SOD and MDA contents was observed in each group. Three hours after injury, MDA level in HHE group began to ascend, reaching the peak level at 6 h that was significantly higher than those in the other 2 groups (P<0.05); SOD level underwent a reverse change, which was significantly lower in HHE group than in the other groups with that in HA group being the highest. Comparable SOD levels in the groups occurred at the time of 14 h, which happened to MDA levels 4 h later. CONCLUSION: Gunshot injuries in the limbs of dogs exposed to hot and humid environment induces increased free oxygen and aggravated the lipid peroxidation, which can be alleviated by heat acclimatization.

Dynamic changes of TNF-alpha content in dogs after gunshot wound in the limbs in hot and humid environment.

OBJECTIVE: To investigate the dynamic changes of TNF-alpha content in the plasma and body tissues of dogs after gunshot wound in the limbs in hot and humid environment. METHODS: Eighteen dogs with gunshot wound were divided into 3 groups (6 in each), one observed in normal environment (Ne group) and the other two in hot and humid environment including a heat-acclimatized group (HA group) and a non-acclimatied group (NHA group). The contents of tumor necrosis factor-alpha (TNF-alpha) in the plasma and muscle tissues from the gunshot wound tract were measured at 0, 1, 3, 4, 6, 8, 10, 14, 18, 24 h respectively after the injury. RESULTS: TNF-alpha content in the muscular tissue from gunshot wound tract was significantly higher than that in the plasma (P<0.05). At 4 h after injury, TNF-alpha content started to rise in NE group, reaching the peak level at 14 h, the rise and peaking of which occurred at 3 and 10 h respectively in NHE group with higher peak level than that in NE group (P<0.05). The changes of TNF-alpha participates in the pathophysiological process after gunshot wound in hot and humid environment and may play the role of and indicator for the pathological changes that take place after the injury.

[The regulatory effect of human bone morphogenetic protein 7 gene transfer on the proliferation and differentiation of rabbit bone marrow mesenchymal stem cells].

OBJECTIVE: To detect the proliferation and differentiation of rabbit bone marrow mesenchymal stem cells (BMSc) transferred by retroviral vector carrying human bone morphogenetic protein 7 (hBMP-7) gene. METHODS: hBMP-7-expressing replication-deficient retroviral vector(PT-PLNCX2-hBMP7) was reconstructed using clone technique and recombinant DNA technique. BMSc were infected with the virus granules. The protein of BMP-7 gene in transferred cells were determined by immunohistochemistry. The proliferativity of the transferred cell were assayed by methabenzthiazuron (MTT) method and flow cytometer. Alkaline phosphatase (ALP) were also detected using enzyme kinetics. RESULTS: Cells transferred by PT-PLNCX2-hBMP7 expressed abundant hBMP7 protein in the cytoplasm. Positive findings were not found in those cells that were not transferred. After infected with virus there were not significant difference of cell proliferation and cell cycle between the cells transferred by hBMP-7 or not (P > 0.05). ALP activity in transferred cells were increased significantly (P < 0.01). CONCLUSIONS: hBMP-7 can be transferred and stably expressed in the cultured rabbit bone marrow stem cells. Proliferation and cell cycle of the transferred cell were not affected. hBMP7 gene transfer can be used to induce differentiation of BMSc into osteoblast-like cells.

[The method of accelerating osteanagenesis and revascularization of tissue engineered bone in big animal in vivo].

OBJECTIVE: To study whether tissue engineered bone can repair the large segment bone defect of large animal or not. To observe what character the fascia flap played during the osteanagenesis and revascularization process of tissue engineered bone. METHODS: 9 Chinese goats were made 2 cm left tibia diaphyseal defect. The repairing effect of the defects was evaluated by ECT, X-ray and histology. 27 goats were divided into three groups: group of CHAP, the defect was filled with coral hydroxyapatite (CHAP); group of tissue engineered bone, the defect was filled with CHAP + bone marrow stroma cells (BMSc); group of fascia flap, the defect was filled with CHAP + BMSc + fascia flap. After finished culturing and inducing the BMSc, CHAP of group of tissue engineered bone and of fascia flap was combined with it. Making fascia flap, different materials as described above were then implanted separately into the defects. Radionuclide bone imaging was used to monitor the revascularization of the implants at 2, 4, 8 weeks after operation. X-ray examination, optical density index of X-ray film, V-G staining of tissue slice of the implants were used at 4, 8, 12 weeks after operation, and the biomechanical character of the specimens were tested at 12 weeks post operation. RESULTS: In the first study, the defect showed no bone regeneration phenomenon. 2 cm tibia defect was an ideal animal model. In the second study, group of CHAP manifested a little trace of bone regeneration, as to group of tissue engineered bone, the defect was almost repaired totally. In group of fascia flap, with the assistance of fascia flap which gave more chance to making implants to get more nutrient, the repair was quite complete. CONCLUSIONS: The model of 2 cm caprine tibia diaphyseal defect cannot be repaired by goat itself and can satisfy the tissue engineering's demands. Tissue engineered bone had good ability to repair large segment tibia defect of goat. Fascia flap can accelerate the revascularization process of tissue engineered bone. And by this way, it augment the ability of tissue engineered bone to repair the large bone defect of goat.

[Effect of bone morphogenetic protein microspheres on biological behavior of rabbit bone marrow stem cells].

OBJECTIVE: To evaluate the effect of bone morphogenetic protein (BMP) on the biological behavior of bone marrow stem cells (BMSCs) of rabbits. METHODS: BMP was either enwrapped or not in the microspheres made of chitosan and sodium alginate, and the biocompatibilities of the composites were examined by means of cell culture. The BMSCs were cultured with the two kinds of microspheres respectively, and the cell extension rate, proliferation, alkaline phosphatase activity and Coomassie blue staining of the cells were assayed. RESULTS: Inhibition of BMSC proliferation did not occur in response to in vitro culture with the microspheres, but alkaline phosphatase activity and D(lambda) values of Coomassie blue staining increased significantly in the cells cultured with BMP microspheres. CONCLUSION: BMP can increase the osteogenic capacity of BMSCs in vitro with the microspheres made of chitosan and sodium alginate as the carrier.

Effect of X-ray irradiation on limb allograft rejection in adult rats.

OBJECTIVE: To establish appropriate animal models for observing the effects of X-ray irradiation on limb allograft rejection. METHODS: Wistar rats were used as donators and SD rats as recipients, the latter divided into 2 groups, namely irradiation group and non-irradiation group according to pretransplant treatment with or without X-ray radiation (5 Gy) on the part of the donators. The donor limbs were transplanted into SD rats who had their own limbs cut off, and the sciatic nerve, femoral nerve, and femoral artery and femoral vein were anastomosed in operation. After the operation, all the recipients were given benzathine benzylpenicillin, and their vital signs, together with changes of the allografts, observed. RESULTS: A total of 16 rats received transplantation that was successful in 6 rats of non-radiation group and 7 of irradiation group. The graft survival averaged 12.0+/-2.4 d in non-irradiation group and 24.7+/-8.1 d in irradiation group, showing significant difference in the lengths of survival between the 2 groups (P<0.05). CONCLUSION: The graft survival time in rats can be significantly prolonged by pretransplant irradiation of the allograft, which also help control acute rejection of the allograft.

[Repair of tibial defect with tissue-engineered bone graft and radionuclide bone imaging in goats].

OBJECTIVE: To observe the effect of tissue-engineered bone grafts in repairing large tibial defect in goats, and assess the value of radionuclide bone imaging in monitoring the therapeutic effect of this approach. METHODS: Tibial defects measuring 2 cm was artificially made in the left tibia of 27 normal goats that were subsequently divided into 3 groups (9 each) to undergo treatment with tissue-engineered bone grafts, artificial bone grafts or without any grafts (as control group) respectively. The tissue-engineered bone grafts contained bone marrow stroma cells (BMSCs) of the goats and coral hydroxyapatite (CHAP), while the artificial bone grafts were from CHAP only. After the operations, radionuclide bone imaging was used to monitor the therapeutic effects at 2, 4 and 8 postoperative weeks. RESULTS: The 99mTc-MDP uptake of the region of interest (ROI) and the target to non-target ratios (T/NT) of the control group did not indicate any process of revascularization or bone regeneration. An increasing tendency of the revascularization and bone regeneration, in contrast, was observed in goats receiving the artificial bone grafts, a tendency that was far more obvious in the goats with tissue-engineered bone grafts. CONCLUSION: Tissue-engineered bone graft is eligible in repairing large defect in the caprine tibia, and radionuclide bone imaging may accurately monitor the revascularization and bone regeneration after the bone graft implantation.

[Biocompatibility of adult human osteoblasts with coral-derived hydroxyapatite in vitro].

OBJECTIVE: To study the biocompatibility of the osteoblasts from adult human bone marrow with coral-derived hydroxyapatite (CHA) in in vitro culture. METHODS: Bone marrow was obtained from healthy adult subjects and cultured in Dulbecco's modified Eagle's medium (DMEM) containing 10 % fetal bovine serum. The subsequent cell passaging was conducted in conditioned medium containing dexamethasone, beta-sodium glycerophosphate and ascorbic acid, with the osteoblasts in culture then divided into CHA group (in which the cells were cultured with CHA) and osteoblasts group (without CHA). The proliferation and differentiation of all the cultured cells were observed at different time points under inverted phase contrast microscope, optical microscope with HE staining and scanning electron microscope respectively. Proliferation of the cultured cells were evaluated by MTT assay, and the activity of alkaline phosphatase and total micro-protein contents in these cultured osteoblasts were quantitatively detected. RESULTS: The osteoblasts from adult human bone marrow grow well in vitro, regardless of the presence of CHA, with biological and morphological characteristics similar to those of normal osteoblasts. CHA improved the adhesion, growth and proliferation of the cultured cells, showing no adverse effects on the cell functions. CONCLUSION: CHA is an optimal scaffold material for bone tissue engineering, which may potentially find clinical application for bone defect repair.

[mRNA expression of recombinant human bone morphogenetic protein 7 in skeletal muscle satellite cells with retroviral vector-mediated rhBMP7 gene transfection].

OBJECTIVE: To detect the expression of recombinant human bone morphogenetic protein7 (rhBMP7) in skeletal muscle satellite cells (SMSCs) with rhBMP7 gene transfection mediated by retroviral vector. METHODS: rhBMP7 gene was reconstructed in retroviral vector and transferred into packaging cells PT67 via liposome reagent, with the positive cell clones selected with G418. In vitro cultured SMSCs were transfected with the virus granules secreted by PT67 cells and followed by G418 selection. Reverse transcriptase-polymerase chain reaction was utilized for analysis for rhBMP7 mRNA in the transfected cells. RESULTS: rhBMP7 retrovirus vector was successfully constructed and transferred into PT67 cells, and abundant mRNA expression of rhBMP7 was observed in the skeletal muscle satellite cells transfected with the virus and selected with G418. CONCLUSION: rhBMP7 gene can be transferred into the skeletal muscle satellite cells via retroviral vector to yield effective rhBMP7 mRNA expression.