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.

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.

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.

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.

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.

Microsurgical techniques used to construct the vascularized and neurotized tissue engineered bone.

The lack of vascularization in the tissue engineered bone results in poor survival and ossification. Tissue engineered bone can be wrapped in the soft tissue flaps which are rich in blood supply to complete the vascularization in vivo by microsurgical technique, and the surface of the bone graft can be invaded with new vascular network. The intrinsic vascularization can be induced via a blood vessel or an arteriovenous loop located centrally in the bone graft by microsurgical technique. The peripheral nerve especially peptidergic nerve has effect on the bone regeneration. The peptidergic nerve can be used to construct the neurotized tissue engineered bone by implanting the nerve fiber into the center of bone graft. Thus, constructing a highly vascularized and neurotized tissue engineered bone according with the theory of biomimetics has become a useful method for repairing the large bone defect. Many researchers have used the microsurgical techniques to enhance the vascularization and neurotization of tissue engineered bone and to get a better osteogenesis effect. This review aims to summarize the microsurgical techniques mostly used to construct the vascularized and neurotized tissue engineered bone.

The potential use of allogeneic platelet-rich plasma for large bone defect treatment: immunogenicity and defect healing efficacy.

Autologous platelet-rich plasma (PRP) has been extensively investigated for large bone defect treatment, but its clinical application is harassed by controversial outcome, due to highly variable PRP quality among patients. Alternatively, allogeneic PRP from well-characterized donors cannot only generate more consistent and reliable therapeutic effect but also avoid harvesting large quantities of blood, an additional health burdens to patients. However, the use of allogeneic PRP for bone defect treatment is generally less investigated, especially for its immunogenicity in such application. Here, we meticulously investigated the immunogenicity of allogeneic PRP and evaluated its healing efficacy for critical-sized defect treatment. Allogeneic PRP contained 4.1-fold and 2.7- to 4.9-fold higher amount of platelets and growth factors than whole blood, respectively. The intramuscular injection of allogeneic PRP to rabbits did not trigger severe and chronic immunoresponse, evidenced by little change in muscular tissue microstructure and CD4⁺/CD8⁺ T lymphocyte subpopulation in peripheral blood. The implantation of allogeneic PRP/deproteinized bone matrix (DPB) constructs (PRP+DPB) successfully bridged 1.5-cm segmental radial defects in rabbits, achieving similar healing capacity as autologous MSC/DPB constructs (MSC+DPB), with greater bone formation (1.1-1.5×, p<0.05) and vascularization (1.3-1.6×, p<0.05) than DPB alone, shown by histomorphometric analysis, bone mineral density measurement, and radionuclide bone imaging. Furthermore, the implantation of both allogeneic PRP- and autologous MSC-mediated DPB constructs (PRP + MSC + DPB) resulted in the most robust bone regeneration (1.2-2.1×, p<0.05) and vascularization (1.3-2.0×, p<0.05) than others (PRP+DPB, MSC+DPB, or DPB alone). This study has demonstrated the promising use of allogeneic PRP for bone defect treatment with negligible immunogenicity, great healing efficacy, potentially more consistent quality, and no additional health burden to patients; additionally, the synergetic enhancing effect found between allogeneic PRP and autologous MSCs may shed a light on developing new therapeutic strategies for large bone defect treatment.

[Three-dimensional reconstruction technique in gastrocnemius flap surgery: initial clinical application].

OBJECTIVE: To discuss the experience with three-dimensional reconstruction technique in initial clinical application in gastrocnemius muscle flap surgery. METHOD: From 2007 to 2008, 7 patients received gastrocnemius muscle flap surgeries to repair the wounds. Preoperative CT angiography or magnetic resonance imaging (MRI) was performed after injection of the contrast media for individualized three-dimensional gastrocnemius muscle flap reconstruction using Amira4.1 software. According to the size of the defect in the wound, individualized three-dimensional gastrocnemius muscle flap was designed and harvested from the posterior leg. RESULTS: Individualized three-dimensional reconstruction of the gastrocnemius flap was performed in 7 cases, and the reconstructed flaps clearly displayed the blood vessels, skin and the adjacent three-dimensional structures. In 6 cases the main perforating branched and trunk of the blood vessels in the designed flap were consistent with the surgical findings; in 1 case, the perforating branches failed to be clearly displayed in the designed flap, and surgical examination identified perforating branches with an average diameter of 0.5 mm (minimally 0.3 mm). The flaps survived in all the 7 cases. CONCLUSIONS: Three-dimensional reconstruction of the gastrocnemius flap based on the lower limb CT angiography or MRI allows three-dimensional observation of the anatomy of the flap and accurate marking of the extent of the flap to be harvested, therefore avoiding intraoperative injuries to the blood vessels to better survival of the flaps.

Replantation of articular composite tissue masses severed from extremities.

SUMMARY: The techniques and outcomes of in situ replantation are discussed for managing 5 cases of articular composite tissue masses severed from an extremity (digit). All 5 cases treated with in situ replantation survived. Rehabilitation was performed after surgery. Follow-ups of 2-5 years showed good appearance, satisfactory functional and sensory recovery. In situ replantation is indicated for an articular composite tissue masses severed from an extremity (digit), if its structure is complete and a blood supply vessel in the mass is available for anastomosis. Replantation can achieve better outcomes than transfer or grafting of adjacent skin or osteocutaneous flaps, or transplantation of a metatarsophalangeal or interphalangeal joint.

Reconstruction of phalangeal articulations of the hand with vascularised phalangeal articulations of foot.

SUMMARY: Since arthroplasty, prosthetic replacement and non-vascularised articulation autografting do not normally produce very satisfactory results for ankylosis of metacarpophalangeal and interphalangeal joints, the authors performed reconstruction of phalangeal articulations of the hand using vascularised phalangeal articulations of the foot in 11 patients with ankylosis of the metacarpophalangeal and interphalangeal joints of hand due by trauma. Procedures included reconstruction of 9 hand metacarpophalangeal joints with vascularised grafting of pedal metatarsophalangeal joints in six patients, reconstruction of the hand metacarpophalangeal joints with grafting of vascularised proximal interphalangeal joints of foot in two patients and reconstruction of the hand proximal interphalangeal joints with grafting of vascularised proximal interphalangeal joints of foot in three patients. Early functional exercise was encouraged in all cases post-operatively. Follow-up ranged from 3 to 10 years and revealed that 9 cases had normal appearance and length of recipient area, 1 had slightly clumsy dorsal skin in the hand and 1 had slight dorsal angulation of a metacarpal bone. Recovery of joint range of motion was satisfactory. Radiographic, gross and sensation examinations also showed good operative outcomes. The authors believe that vascularised pedal metatarsophalangeal joints, with a rich blood supply, can be grafted to effectively reconstruct the finger joints with good function. A low rate of degeneration results because pedal and hand metatarsophalangeal joints are similar in anatomy and physiological function.

[Green fluorescent protein as a tracer of bone marrow stromal cells in bone tissue engineering in rhesus].

OBJECTIVE: To observe the role of green fluorescent protein (GFP) in tracing rhesus bone marrow stromal cells (rBMSCs) during tissue-engineered bone formation in vivo. METHODS: Ad5.CMV-GFP was amplified by infecting QBI-293A cells, and the bone marrow was harvested from the ilium of adult male rhesus to obtain rBMSCs, which were cultured and passaged in vitro. GFP was transfected into the third-passage rBMSCs via adenovirus vector and the labeled cells were inoculated into absorbable HA scaffold and cultured for 3 days, with untransfected rBMSCs as control, before the cell-matrix compounds were implanted into the latissimus dorsi muscles of rhesus. Samples were harvested at 6 week and embedded in paraform, and ground sections of the bone tissue were prepared to observe green fluorescence under laser scanning confocal microscope. Propidium iodide staining of the sections was also performed for observation. RESULTS: The rBMSCs grew well after GFP transfection, and green fluorescence could be seen 24 h after the transfection and became stronger till 48 h, with a positive transfection rate beyond 80%. Six weeks after cell implantation, the rBMSCs labeled by GFP-emitted green fluorescence were detected in the bone tissue under laser scanning confocal microscope. CONCLUSION: GFP can effectively trace BMSCs during bone tissue engineering, and the transplanted BMSCs constitute the main source of bone-forming cells in bone tissue engineering.

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.

[Perfusion-weighted magnetic resonance imaging for monitoring vascularization in tissue-engineered bone in rhesuses].

OBJECTIVE: To assess the value of perfusion-weighted magnetic resonance (MR) imaging (PWMRI) in monitoring vascularization in tissue-engineered bone graft. METHODS: Tibial diaphyseal defect of 20 mm was induced in 25 lower limbs of 13 rhesuses and fixed with an AO reconstruction plate with 7 holes. The monkeys were randomized into 5 groups according to the materials used for defect filling: group A, with beta-tricalcium phosphate (beta-TCP), bone marrow stromal cells (BMSCs) and blood vessel bundles; group B, with beta-TCP and blood vessel bundles; group C, with beta-TCP and BMSCs; group D, with beta-TCP, and group E without filling. PWMRI, X-ray, and radionuclide imaging were carried out at weeks 4, 8, 12 postoperatively. The maximum slope rates of the single intensity-time curve (SS(max)) and the baseline values (SI(baseline)) on the same time points were calculated. Transmittances on the X-ray films and isotope counts in the region of interest (ROI) were assessed and calculated. RESULTS: Compared with other groups, group A showed the highest SS(max) at weeks 4, 8, and 12 postoperatively, and its SS(max) at week 8 was significantly higher than that at week 4 (P=0.003). The SS(max) was positively related to isotope counts in ROI at week 8 after operation (r(s)=0.899, P=0.038), and inversely related to transmittance on X-ray films at week 12 (r(s)=-0.892, P=0.042). CONCLUSION: The SS(max) of the single intensity-time curve can accurately reflect the vascularization of the tissue-engineered bone graft, and PWMRI allows sensitive, quantitative, noninvasive and radiation-free vascularization monitoring.

[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.

[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.

[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.

[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.