Objective To investigate application feasibility of the heterogeneous de protein cancellous bone in the treatment of lumber intertransverse process spinal fusion.Methods Thirty 6 ～ 8 month healthy male goats were selected to make intertransverse bone bed,and were randomly divided into groups Ⅰ and Ⅱ,with 15 in each group.Group Ⅰ was the left side implanted the tissue engineering bone (A group),and the right side implanted recombinant human bone morphogenetic protein-2 (rhBMP-2) composite bone (B group);Group Ⅱ was the left implanted autologous bone (C group),and the right side implanted simple heterogeneous de protein cancellous bone (D group).Each group was observed with the Xray,histological morphology,and maximum bending load.Results (1) In groups B and D,postoperative 4-,8-and 12-week X-ray scores were lower than that of group C (P ＜0.05);In group A,postoperative 4-,8-and 12-week X-ray score were (5.30 ± 0.41) points,(8.54 ± 0.89) points,and (11.25 ± 0.91)points,compared to group C,without statistically significant difference (P ＞ 0.05);(2) For groups A,B and D,postoperative 4-and 8-week histological scores were significantly lower than those in group C (P ＜0.05).For group A,postoperative 4-and 8-week histological scores were significantly higher than those of groups B and D (P ＜0.05).The histological scores of groups B and D postoperative 12 weeks were lower than those of group C (P ＜0.05).The histological score of group A postoperative 12 weeks was (11.04 ± 1.52) points,compared to group C without statistically significant difference (P ＞ 0.05);(3) The maximum bending load of groups B and D was lower than that of group C (P ＜ 0.05).The maximum bending load of group A was (60.32 ± 10.10) N,compared to group C without statistically significant difference (P ＞ 0.05).Conclusions The construction of tissue engineered bone by the heterogeneous de protein cancellous bone,its osteogenesis is close to autogenous bone,but simple heterogeneous de protein cancellous bone osteogenesis is poor.

Objective To explore the diagnosis and operative treatment of spinal tumor. Methods Clinical symptom, diagnosis and operative treatment of 22 patients with spinal tumor in our institution from August 1987 to August 2002 were analysed retrospectively. Results Combining with patients' clinical symptoms and results of corresponding examination, all cases were diagnosed before operation. After operation, all the patients recovered with different outcome except 3 patients died. There were 18 cases nearly cured. Conclusion The clinical symptoms of the patients with spinal tumor have some specialty. The preoperative diagnosis mainly depends on MRI. The various operative modes should be taken according to tumor localization.

Objective:To investigate the feasibility of construction of tissue engineered cartilage by co-culture of bone marrow mesenchymal stem cells (BMSCs) and costal chondrocytes (CCs),and to provide theoretical basis and experimental basis for clinical repair of articular cartilage defects by Wuzhishan miniature pig knee cartilage defects with co-cultured cells.Methods:Density gradient centrifugation method was used to isolate BMSCs from Wuzhishan miniature pig.The double enzyme digestion method was used to isolate CCs.The passage 3 generation of BMSCs and passage 2 generation of CCs were randomly divided into 3 groups:a co-culture group of BMSCs∶CCs for 1∶2 (Group A),a simple CCs (Group B),and a simple BMSCs (Group C).The cell growth curve was drawn,and the content of glycosaminoglycan (GAG) of external separation in chondrocytes was determined.The 12 Wuzhishan miniature pigs were randomly divided into a co-culture cells/collagen membrane experimental group,a collagen membrane control group and the blank group.In the co-culture cells/collagen membrane experimental group,the co-cultured cells/collagen membrane were implanted into the cartilage defects of the mandibular condyle;in the collagen membrane control group,only collagen membrane was implanted;while in the blank group,nothing was implanted.Six animals were sacrificed at 8 and 16 weeks after surgery respectively (2 animals in each group).General observation,cartilage histological score and histopathological examination were carried out.Results:The BMSCs and co-culture cells grew well.The biological activity of CCs was good.After 16 weeks of operation,the repair tissues in the co-cultured cells/collagen membrane experimental group showed hyaline cartilage features:smooth,flat,and integrated well with the surrounding cartilage and subchondral bone.The collagen membrane in the collagen membrane control group was fibrously repaired.Repair tissue gross score in the co-culture cells/collagen membrane experimental group was significantly better than that in the collagen membrane control group and the blank group (both P<0.05),but there was no significant difference between the collagen membrane control group and the blank group (P>0.05).Conclusion:BMSCs,CCs and co-cultured cells can function as the seed cells for cartilage tissue engineering,and the co-culture cells (BMSCs∶CCs=1∶2) possess more advantages;the short-term effect of co-culture cells with collagen membrane on repairing cartilage defects is satisfied.

Objective To investigate the association of endothelial nitric oxide synthase (eNOS) polymorphisms and avascular necrosis of femoral head (ANFH),to explore possible relationship between the ANFH incidence and 27-bp repeat polymorphism in intron 4 and G894T polymorphism in exon 7.Methods Totally 125 atraumatic ANFH patients and 126 healthy controls without hip trauma history were enrolled.Gene polymorphisms in 27-bp repeat polymorphism in intron 4 and G894T polymorphism in exon 7 were determined.Medical history was collected for etiology analysis.Results In between-group comparison,the frequency of b/a genotype intron 4 in ANFH group was significantly higher than that in healthy control group [19/125 (15.2％) vs 6/126 (4.8％),P =0.0001,OR =4.501],and the result of G/T genotype exon 7 in the ANFH group also indicated a statistical significance with the healthy control group [30/125 (24.0％) vs 17/126 (13.5％),P =0.0094,OR =3.804].In subgroup analysis,genotype b/a and G/T were especially higher in the idiopathic group than that in the healthy control group.Conclusions eNOS gene polymorphisms might be a risk factor of ANFH,there is underlying relevance of eNOS and the disease.Both 27-bp repeat polymorphism in intron 4 and G894T polymorphism in exon 7 are associated with ANFH incidence.

Objective To observe the short-term effect of radial head replacement for the treatment of terrible triad of the elbow.Methods In the period between June 2011 and June 2012,the radial head replacements were carried out in six patients with terrible triad of elbow.There were five acute elbow fracture-dislocation cases and one old fractnre case.All cases underwent open reduction and fixation coronoid fracture with screws or nonabsorbable sutures,radial head replacement,repair of lateral ligament complex and repair or reconstruction of annular ligament.All patients initiated the rehabilitation program under supervision within 5-7 days after surgery.The X-ray and Computed tomography of elbow was used to evaluate the posi tion of radial head prosthesis.Functional outcome of elbow was assessed by Mayo Elbow Performance Score (MEPS) and complications.Results All patients were participated in follow-up for 10-24 months and the mean duration of follow-up was 16.8 months.The MEPS was from 85 to 95 points and the mean MEPS was 91.7 points.The outcome of MEPS was excellent in 5 cases,good in 1 case.The range motion of elbow flexion was from 82 to 95 degrees and the average of elbow flexion was 87 degrees; the range motion of extension was from 15 to 32 degrees and the average of elbow extension was 21 degrees.The range motion of fore-ann pronation was from 82 to 90 degrees with 86 degrees in average.The motion range of supination was from 45 to 80 degrees with 56 degrees in average.All cases got persistent stability in elbow with painless movement.None of patient suffered traumatic arthritis and infection.The motion range of elbow was restricted due to the unfit relative position of radial head prosthesis and capi tellum in one case,characterized by decreasing radiocapitellar joint space.Two cases developed into heterotopic ossification in elbow without hampering the motion range of elbow.Conclusion The outcome of the radial head replacement on the treatment of radial head fracture in the elbow terrible triad was satisfactory.However,the longer duration of observation was essential to verify the favorable effect of radial head replacement on dealing with terrible triad in elbow.

Tendon tissue engineering is a novel therapeutic strategy for severe tendon injury and loss. Adipose derived stem cells (ASCs) have been studied extensively, due to their potency to differentiate into musculoskeletal tissue precursors such as osteoblasts, chondrocytes, adipocytes, and tendocytes under specific cues and high ability of proliferation. Resources of ASCs are ubiquitous and isolation of ASCs is secure, simple and minimally invasive. Mounting evidences demonstrate that ASCs may be involved in tendon tissue engineering and repair the severe injury of tendon under stimulation of various growth factors and other appropriate fittings.
Adipocytes ; Adipose Tissue ; cytology ; Cell Differentiation ; Chondrocytes ; Humans ; Osteoblasts ; Stem Cells ; cytology ; Tendon Injuries ; therapy ; Tendons ; cytology ; Tissue Engineering ; Wound Healing

OBJECTIVE: To determine the effect of bone cement, with different amounts of alendronate on osteoblast, and determine the cytotoxicity of alendronate-integrated bone cement from the viewpoint of cell biology. METHODS: According to different additions (0, 10, 50, 100, 500, 1 000 mg) of alendronate in 50 g Cemex®XL bone cement powder, the experiments were divided into 6 groups, namely G0-G5 groups. In all groups, the adhesive capacity of osteoblast-like cells MG-63 was evaluated by electron microscope, the optical density (OD) value of cells by MTT colorimetry method, the alkaline phosphatase activity (AKP) by AKP assay kit, the apoptosis rates by Annexin-V-FITC apoptosis detection kit, and the bone mineralization potentiality by phase contrast microscope. RESULTS: The adhesive capacity of MG-63 was good in all groups. Compared with the G0 group, the cell apoptosis was inhibited in G1-G4 groups while in G5 group the cell apoptosis was promoted and cell proliferation was inhibited (P<0.05). In all groups, no significant difference was found in alkaline phosphatase activity and bone mineralization potentiality (P>0.05). CONCLUSION Less than 500 mg alendronate added in Cemex®XL 50 g bone cement powder has no cytotoxicity on osteoblasts.
Alendronate ; Apoptosis ; Bone Cements ; Cell Adhesion ; Cell Proliferation ; Humans ; Osteoblasts ; cytology ; drug effects

OBJECTIVE: To determine the effect of alendronate amount on the static mechanical properties of bone cement before elution and 4 weeks after elution, and determine suitable amount of alendronate in cement from the perspective of biomechanics. METHODS: Samples of compression strength and bending strength (modulus) were prepared with bone cement adding 0, 10, 50, 100, 500 and 1000 mg alendronate in 50 g Cemex®XL bone cement powders respectively (named as G0-G5 groups). The compression strength, bending strength, and bending modulus of bone cement were examinated by INSTRON 8032 tester before the elution and 4 weeks after the elution. Some broken samples of 4-point bending test were examinated by scanning electronic microscopy (SEM), and some other samples of bending strength (modulus) tests by micro-CT. RESULTS: No significant difference was found in the compression strength before the elution,4 weeks after elution in the 6 groups, and before and after the elution in the respective groups (P>0.05). Compared with G0 group, G1-G4 groups had no obvious difference in the bending strength before the elution and 4 weeks after the elution (P>0.05), while G5 group had difference (P<0.05). There was no significant difference in the bending strength before and after the elution in the respective groups (P>0.05). Taken G0 group as the control group, G1-G4 groups had no visible effects on the bending modulus 4 weeks after the elution (P>0.05), while G5 group decreased it significantly (P<0.05). Bending modulus before the elution in the 6 groups didn't show obvious difference (P>0.05), while bending modulus before and after the elution in the respective groups displayed a marked difference (P<0.05). CONCLUSION Less than 500 mg alendronate added in Cemex?XL 50 g bone cement powder does not decrease the compressive strength, flexural strength and flexural modulus before the elution and 4 weeks after the elution.
Alendronate ; chemistry ; Bone Cements ; Compressive Strength ; Elastic Modulus ; Materials Testing ; Microscopy, Electron, Scanning ; Powders

Objective This study is to compare flap-viability-related complications, coverage reach, recon-struction outcomes and donor-mobidities between distally-based peroneal artery perforator-plus fasciocutaneous (DPAPF)flap and distally-based posterior tibial artery perforator-plus fasciocutaneous(DPTAPF)flap for recon-struction of soft-tissue defects over the distal lower leg, ankle and foot, and thus provide evidence for selection of the flaps. Methods Between April, 2002 and February, 2012, 216 and 59 patients underwent the reconstructions with DPAPF flaps(peroneal group)and DPTAPF flaps(posterior tibial group)respectively. We subdivided the distal lower leg, ankle and foot into 12 subregions. In all the patients, flap-viability-related complications and its potential risk factors(including age,sex,etiology,location of top edge,location of pivot point,length and width of both the skin is-land and adipofascial pedicle, length-width ratio, and total length), coverage reach(the subregion in which the most distal part of the reconstructed defect lies),duration of flap elevation and hospital stay were compared between the two groups. In patients with at least 3 months postoperative follow-up, comparative study of reconstruction outcomes, pa-tient's satisfaction with flap appearance and donor-site morbidities were performed between the groups. Results Partial necrosis rate in the peroneal of the posterior tibial group were 12.0 percent versus 20.3 percent,respectively(P> 0.05). Marginal necrosis and overall complication (including partial and marginal necrosis)rates in the peroneal group(1.9 percent and 13.9 percent, respectively)were significantly lower than those in the posterior tibial group (8.5 percent and 28.8 percent,respectively)(P<0.05).Incidence of partial necrosis of the flaps for the defects over subregions 7 to 10 in the peroneal group(7 of 41)was significantly lower than that in the posterior tibial group(2 of 2).There was no difference in reconstruction outcomes and patient's satisfaction with flap appearance in both groups(P >0.05).Incidences of hypertrophic scar,itching and pigmentation at the donor site were significantly lower in the peroneal group(P<0.05). Conclusion DPAPF flap is superior to DPTAPF flap in reliability,safe coverage reach and less donor-site morbidities.The former is recommended as the first choice when local pedicle flaps are considered to recon-struct soft-tissue defects over the distal lower leg,ankle and foot.

Objective To investigate the relationship between vascular endothelial cell nitric oxide synthase (eNOS) gene polymorphism and the pathogenesis of avascular femoral head necrosis (ANFH).Methods The eNOS full-length CDS fragments,containing 894G or 894T separately,was subcloned into lentiviral expression vector,and then infect the human umbilical vein endothelial cells (HUVEC).The contents of nitric oxide NO and cyclic guanosine monophosphate (cGMP) in cell culture supernatant were detected in a time-dependent manner.The luciferase-labeled reporter plasmid pNF-κB-luc was co-transfected with the reference control plasmid pRL-TK into HUVEC cells infected with LV-eNOS-894G,LV-eNOS-894T,and control lentivirus (LV-NC) for 48 h.The luciferase activity of each group was detected.The expression of nuclear factor (NF)-κB protein and eNOS protein in HUVEC cells were detected by Western blot assay.The HUVEC cells in each group were co-cultured with hFOB1.19 cells,and the concentration of alkaline phosphatase (ALP) or osteocalcin (OCN) in the supernatant were detected at different time points.Results The contents of NO and cGMP in the cell culture supernatant of the full-length lentivirus expressing eNOS gene (containing 894G or 894T) were significantly higher than that in the empty cell group and the empty vector group,and the contents of NO and cGMP in the cell culture supernatant of the 894G group were significantly higher than that of 894T group (P ＜ 0.01).Compared with blank cells,the expression levels of NF-κB/p65 protein and eNOS protein were significantly increased in cells expressing eNOS,and the expression of NF-κB/p65 protein in 894G group was significantly increased than 894T group,but there was no difference in eNOS protein expression between the 894G and 894T groups;each group of HUVEC cells were co-cultured with hFOB1.19 osteoblasts,and at each of the same time points,the concentrations of ALP and OCN in the cell culture supernatant expressing lentivirus were significantly higher than that in the empty cell group and the empty vector group,and the ALP and OCN concentrations in the cell culture supernatant of the 894G group were significantly higher than those in the 894T group.Conclusions The eNOS gene exon G894T mutation reduces the levels of nitric oxide and cGMP produced by vascular endothelial cells through the eNOS-NO pathway,affecting the expression levels of NF-κB/p65 protein and eNOS protein,and reducing osteoblast activity,and blood supply to blood vessels.It may be one of the pathogenic mechanisms leading to femoral head necrosis.