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1.
Orthop Surg ; 14(5): 902-910, 2022 May.
Artigo em Inglês | MEDLINE | ID: mdl-35441498

RESUMO

OBJECTIVE: To investigate the application of impaction bone grafting (IBG) combined with Ti-alloy mesh for acetabular bone defect reconstruction in total hip arthroplasty (THA) revision and follow up the clinical outcomes and imaging findings. METHODS: The clinical and imaging data of patients who were admitted to our hospital from January 2000 to December 2020 and underwent acetabular bone defects reconstruction using IBG combined with titanium mesh were retrospectively analyzed. Preoperative and post-revision Oxford and Harris scores, and post-revision complications were evaluated. Radiographs were used to determine center of rotation (COR) of the hip joint, transparency line, bone graft fusion, and bone mineral density (BMD) around the hip joint. RESULTS: Significant improvement was observed in both Oxford and Harris scores (P < 0.05). The radiographs taken at the last follow-up examination showed no significant differences in the acetabulum COR, offsets, inclination angle, mean ratio of vertical value, and BMD analysis between the post-revision side and contralateral side (P > 0.05). The follow-up data showed restoration of the mesh implant and graft bone fusion. CONCLUSIONS: The application of IBG combined with titanium-alloy mesh in revision THA patients with acetabular defects was found to provide satisfactory outcomes. However, large-scale studies are still needed to further elucidate the long-term outcomes.


Assuntos
Artroplastia de Quadril , Prótese de Quadril , Acetábulo/cirurgia , Ligas , Artroplastia de Quadril/métodos , Transplante Ósseo/métodos , Seguimentos , Humanos , Falha de Prótese , Reoperação/métodos , Estudos Retrospectivos , Telas Cirúrgicas , Titânio
2.
Medicine (Baltimore) ; 99(36): e22028, 2020 Sep 04.
Artigo em Inglês | MEDLINE | ID: mdl-32899057

RESUMO

Comparison of different anticoagulants in blood management and complications with tranexamic acid (TXA) in total hip arthroplasty (THA) is unclear. Our aim was to compare the efficacy and safety among receiving nadroparin calcium, enoxaparin sodium or rivaroxaban after TXA in THA.150 patients undergoing primary unilateral THA were received 15 mg/kg intravenous TXA (IV-TXA) before skin incision, followed by 1 of nadroparin calcium (Group A), enoxaparin sodium (Group B), or rivaroxaban (Group C) randomly during hospitalization. The primary outcome was hidden blood loss (HBL). Other outcomes such as the maximum hemoglobin (Hb) drop, total blood loss (TBL), the volume of drainage, transfusion rate, length of hospital stay (LOS), and complications were also compared.There were no statistically significant differences in HBL, the maximum hemoglobin (Hb) drop, transfusion rate, and complications among 3 groups. LOS was significantly higher for patients in Group B than Group A (P = .026). Neither deep venous thrombosis (DVT) nor pulmonary embolism (PE) occurred in any group.There were no differences in efficacy and safety in patients undergoing THA receiving nadroparin calcium, enoxaparin sodium, or rivaroxaban after anti-fibrinolysis with TXA.


Assuntos
Anticoagulantes/efeitos adversos , Antifibrinolíticos/efeitos adversos , Perda Sanguínea Cirúrgica/prevenção & controle , Ácido Tranexâmico/efeitos adversos , Administração Intravenosa , Idoso , Anticoagulantes/administração & dosagem , Antifibrinolíticos/administração & dosagem , Artroplastia de Quadril/efeitos adversos , Transfusão de Sangue/estatística & dados numéricos , China/epidemiologia , Enoxaparina/administração & dosagem , Enoxaparina/efeitos adversos , Feminino , Hemoglobinas/análise , Hospitalização , Humanos , Tempo de Internação/tendências , Masculino , Pessoa de Meia-Idade , Nadroparina/administração & dosagem , Nadroparina/efeitos adversos , Sangue Oculto , Estudos Retrospectivos , Rivaroxabana/administração & dosagem , Rivaroxabana/efeitos adversos , Segurança , Ácido Tranexâmico/administração & dosagem , Resultado do Tratamento
3.
Orthop Surg ; 11(4): 653-663, 2019 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-31456320

RESUMO

OBJECTIVE: To investigate the influence of preoperative osteopenia/osteoporosis on periprosthetic bone loss after total hip arthroplasty (THA) and the efficiency of zoledronate (ZOL) treatment in periprosthetic bone preservation. METHODS: This multicenter, prospective cohort study was conducted in four centers between April 2015 and October 2017. Patients were assigned to Normal BMD, Osteopenia, and Osteoporosis+ZOL groups. Patients with osteopenia received daily oral calcium (600 mg/d) and vitamin D (0.5 µg/d), while patients in the Osteoporosis+ZOL group received additional ZOL annually (5 mg/year). Periprosthetic bone mineral density (BMD) in seven Gruen zones, radiographic parameters, Harris hip score, EuroQol 5-Dimensions (EQ-5D) score, and BMD in hip and spine were measured within 7 days, 3 months, 12 months postoperation and annually thereafter. RESULTS: A total of 266 patients were enrolled, while 81 patients that completed the first year follow-up were involved in the statistical analysis. The mean follow-up time was 1.3 years. There were significant decreases of mean BMD in total Gruen zones (-4.55%, P < 0.05) and Gruen zone 1 (-10.22%, P < 0.01) in patients with osteopenia during the first postoperative year. Patients in the Osteoporosis+ZOL group experienced a marked increase in BMD in Gruen zone 1 (+16%) at the first postoperative year, which had a significant difference when compared with the Normal BMD group (P < 0.05) and the Osteopenia Group (P < 0.001). Low preoperative BMD in hip and spine was predictive of bone loss in Gruen zone 1 at 12 months after THA in patients with normal BMD (R2 = 0.40, P < 0.05). CONCLUSIONS: Patients with osteopenia are prone to higher bone loss in the proximal femur after cementless total hip arthroplasty (THA). ZOL, not solely calcium and vitamin D, could prevent the accelerated periprosthetic bone loss after THA in patients with osteopenia and osteoporosis.


Assuntos
Artroplastia de Quadril/efeitos adversos , Conservadores da Densidade Óssea/uso terapêutico , Densidade Óssea/efeitos dos fármacos , Doenças Ósseas Metabólicas/tratamento farmacológico , Osteoporose/tratamento farmacológico , Ácido Zoledrônico/uso terapêutico , Adulto , Idoso , Cálcio , Progressão da Doença , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Estudos Prospectivos , Vitamina D
4.
Indian J Orthop ; 48(4): 347-53, 2014 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-25143636

RESUMO

BACKGROUND: To test whether autologous transplantation of bone marrow-derived mesenchymal stem cells (BM-MSCs) expressing human bone morphogenic protein-2 (hBMP-2) can produce bone in rabbit leg muscles. MATERIALS AND METHODS: MSCs were isolated from BM of the iliac crest of rabbits and then infected with lentiviral vectors (LVs) bearing hBMP-2 and green fluorescent protein under the control of the cytomegalovirus (immediate early promoter). Differentiation of transduced MSCs to osteoblasts in vitro was evaluated with an alkaline phosphatase activity assay and immuohistochemistry against osteoblast specific markers. MSCs expressing hBMP-2 were placed in an absorbable gelatin sponge, which was then transplanted into the gastrocnemius of rabbits from which MSCs were isolated. Bone formation was examined by X-ray and histological analysis. RESULTS: LVs efficiently mediated hBMP-2 gene expression in rabbit BM-MSCs. Ectopic expression of hBMP in these MSCs induced osteoblastic differentiation in vitro. Bone was formed after the MSCs expressing hBMP-2 were transplanted into rabbit muscles. CONCLUSION: Ectopic expression of hBMP-2 in rabbit MSCs induces them to differentiate into osteoblasts in vitro and to form a bone in vivo.

5.
Eur J Orthop Surg Traumatol ; 23(1): 73-9, 2013 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-23412411

RESUMO

OBJECTIVE: Avascular necrosis of the femoral head is one of the most frequently reported complications in patients with systemic lupus erythematosus (SLE) and often requires total hip arthroplasty (THA). Our objective was to analyze the perioperative management, technical problems, clinical outcomes, and complications associated with THA in patients with SLE. METHODS: A total of 28 total hip arthroplasties performed for 24 patients with SLE, including 19 women and 5 men with a mean age of 38.8 years performed from 1998 to 2011 were retrospectively reviewed. SLE disease activity index and ASA class were evaluated preoperatively. WOMAC, HHS, and SF-36 scores were also evaluated in all cases pre- and post-operatively for functional recovery of the hip and health-related quality of life (HRQOL). RESULTS: The average SLE disease activity index was 3.5 points. Three patients were in ASA class I, 12 class II, and 9 were class III (37.5%). The average duration of follow-up was 67.5 months. None of the patients required a revision, and 3 patients died during the follow-up period. A statistically significant improvement in all scores was found comparing pre- and post-operative conditions (P < 0.001). The complication rate was 11.1% with 2 wound infections and 1 urinary tract infection. CONCLUSION: THA is an acceptable method for achieving functional recovery and increasing HRQOL in patients with SLE and ANFH who receive proper perioperative management.


Assuntos
Artroplastia de Quadril/efeitos adversos , Necrose da Cabeça do Fêmur/cirurgia , Lúpus Eritematoso Sistêmico/complicações , Lúpus Eritematoso Sistêmico/cirurgia , Adulto , Idoso , Artroplastia de Quadril/métodos , Artroplastia de Quadril/reabilitação , Feminino , Necrose da Cabeça do Fêmur/complicações , Necrose da Cabeça do Fêmur/diagnóstico por imagem , Seguimentos , Humanos , Masculino , Pessoa de Meia-Idade , Osteoporose/complicações , Osteoporose/cirurgia , Período Pós-Operatório , Qualidade de Vida , Radiografia , Recuperação de Função Fisiológica , Estudos Retrospectivos , Resultado do Tratamento , Adulto Jovem
6.
Eur J Orthop Surg Traumatol ; 23(2): 219-24, 2013 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-23412455

RESUMO

BACKGROUND: Many previous studies have focused on the postoperative complication of postoperative knee pain, infection, knee prosthesis loosening, periprosthetic fractures, and so on. There have been few studies focused on postoperative ecchymosis formation surrounding the wound of the TKA site. A certain degree of effect on the early functional recovery of the patients may occur due to the mental stress caused by the ecchymosis, which raises doubts regarding the success of the surgery. Therefore, it is particularly important to understand the risk factors for postsurgical ecchymosis formation after TKA, and specific measures for preventing ecchymosis should be taken. In this study, we reviewed the record of patients who received TKAs in our hospital, and a comprehensive analysis and assessment was conducted regarding 15 clinical factors causing postsurgical ecchymosis formation. METHODS: The records of 102 patients who received unilateral TKAs between January 2007 and May 2010 were retrospectively analyzed. Patients were divided into two groups based on the occurrence of ecchymosis. RESULTS: Of the 102 patients, 14 (13.7%) developed ecchymosis. Blood transfusion and drainage catheter clamping during the first few postoperative hours had a significant impact on the development of ecchymosis (p < 0.05). There was no difference in age, BMI, operation time, pre- and postoperative platelet count, and length of postoperative anticoagulant therapy between the two groups. Multivariate logistic regression revealed major risk factors for ecchymosis were postoperative blood transfusion (odds ratio (OR) = 15.624) and drainage catheter clamping (OR 14.237) (both, p < 0.05). CONCLUSION: Blood transfusion and drainage catheter clamping after TKA due to excessive blood suction were associated with higher risks for ecchymosis formation surrounding the surgical site.


Assuntos
Artroplastia do Joelho/efeitos adversos , Drenagem/efeitos adversos , Equimose/etiologia , Complicações Pós-Operatórias/etiologia , Reação Transfusional , Idoso , Artroplastia do Joelho/métodos , Catéteres/efeitos adversos , Drenagem/métodos , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Recuperação de Função Fisiológica , Estudos Retrospectivos
7.
J Orthop Res ; 30(10): 1570-6, 2012 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-22488880

RESUMO

To investigate whether platelet-rich plasma (PRP) when used with allograft bone improves the management outcome of displaced intra-articular calcaneal fractures. Over a 7-year period, all displaced type III calcaneal fractures admitted in our department (276 fractures in 254 patients) were randomly divided into three groups according to the plan of management: autograft alone (n = 101), allograft combined with PRP (n = 85), or allograft alone (n = 90). Radiographic imaging and three-dimensional computed tomography were used to assess the thalamic portion, Bohler's angle, the crucial angle of Gissane, and the height, width and length of the calcaneum. The American Orthopedic Foot and Ankle Society (AOFAS) ankle-hind-foot scoring system was used to evaluate the hind foot function at 12, 24, and 72 months postsurgery. At 12 months no significant difference existed in outcome amongst the treatment groups (p > 0.05). However, at 24 and 72 months the results of the autograft, and the allograft combined with PRP, were similar and both were significantly better than that of the allograft alone (p < 0.05). PRP augmented the favorable outcome of allografts in the management of displaced calcaneal fractures, and matched that of autograft used alone. The findings of this study thus support the clinical use of PRP in conjunction with allograft in the treatment of displaced intra-articular calcaneal fractures.


Assuntos
Transplante Ósseo , Calcâneo/lesões , Consolidação da Fratura , Fraturas Intra-Articulares/terapia , Plasma Rico em Plaquetas , Adolescente , Adulto , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Estudos Prospectivos , Recuperação de Função Fisiológica , Adulto Jovem
8.
J Cell Biochem ; 113(3): 888-98, 2012 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-22371969

RESUMO

Human adipose-derived stem cells (hADSC) are capable of differentiating into an osteogenic lineage. It is believed that microRNAs (miRNAs) play important roles in regulating this osteogenic differentiation of human adipose-derived cells, although its molecular mechanism remains unclear. We investigated the miRNA expression profile during osteogenic differentiation of hADSCs, and assessed the roles of involved miRNAs during the osteogenic differentiation. We obtained and cultured human adipose-derived stems cells from donors who underwent elective liposuction or other abdominal surgery at our institution. miRNA expression profiles pre- and post-osteogenic induction were obtained using microarray essay, and differently expressed miRNAs were verified using quantitative real-time polymerase chain reaction (qRT-PCR). The expression of osteogenic proteins was detected using an enzyme-linked immunosorbent assay. Putative targets of the miRNAs were predicted using online software MiRanda, TargetScan, and miRBase. Eight miRNAs were found differently expressed pre- and post-osteogenic induction, among which four miRNAs (miR-17, miR-20a, miR-20b, and miR-106a) were up-regulated and four miRNAs (miR-31, miR-125a-5p, miR-125b, and miR-193a) were down-regulated. qRT-PCR analysis further confirmed the results. Predicted target genes of the differentially expressed miRNAs based on the overlap from three public prediction algorithms: MiRanda, TargetScan, and miRBase Target have the known functions of regulating stem cell osteogenic differentiation, self-renewal, signal transduction, and cell cycle control. We identified a group of miRNAs that may play important roles in regulating hADSC cell differentiation toward an osteoblast lineage. Further study of these miRNAs may elucidate the mechanism of hADSC differentiation into adipose tissue, and thus provide basis for tissue engineering.


Assuntos
Tecido Adiposo/citologia , MicroRNAs/metabolismo , Osteogênese/genética , Células-Tronco/citologia , Células-Tronco/metabolismo , Adulto , Diferenciação Celular/genética , Células Cultivadas , Citometria de Fluxo , Perfilação da Expressão Gênica , Humanos , Pessoa de Meia-Idade , Análise de Sequência com Séries de Oligonucleotídeos , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Adulto Jovem
9.
J Orthop Sci ; 17(3): 219-25, 2012 May.
Artigo em Inglês | MEDLINE | ID: mdl-22323012

RESUMO

BACKGROUND: The effect of body mass index (BMI) on the outcome of total hip arthroplasty (THA) remains controversial. The purpose of this study was to examine whether revision rate and postoperative outcomes following THA were influenced by BMI. MATERIALS AND METHODS: We retrospectively evaluated 714 patients (751 hips) who underwent primary THA in our department between March 1991 and April 2006. They were followed prospectively for 5-20 years with 24 deaths (24 hips) and 33 losses (34 hips). Patients were separated into three groups based on BMI: underweight, normal and obese groups. A survival analysis was performed using revision as the endpoint, and a case-matched study that was matched for age, gender, and laterality was designed; outcomes were assessed with the Harris Hip score, 36-item short-form health survey, complication rate and radiological examination. RESULTS: The preoperative scores were lower for the obese group, and the postoperative scores were higher for the normal group. Patients in the obese group obtained the greatest overall improvement in clinical scores from admission to the last follow-up. We found a significantly higher complication rate in the obese group and underweight group. It appears that being underweight was associated with an increased dislocation rate, and obese patients were more likely to have osteolysis, deep vein embolism, and pulmonary thrombosis. The log rank test for survival showed no significant differences among the three groups. CONCLUSIONS: Abnormal BMI does not prevent functional rehabilitation after THA; however, patients with abnormal BMI have to face higher complication rates and poorer clinical outcomes following this operation.


Assuntos
Artroplastia de Quadril , Índice de Massa Corporal , Qualidade de Vida , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Complicações Pós-Operatórias/epidemiologia , Estudos Retrospectivos , Resultado do Tratamento
10.
Zhonghua Wai Ke Za Zhi ; 48(14): 1069-73, 2010 Jul 15.
Artigo em Chinês | MEDLINE | ID: mdl-21055110

RESUMO

OBJECTIVES: To analyze the reason of revisions no more than 5 years after primary hip replacement, and to discuss the methods how to prevent and manage. METHODS: Retrospectively review 11 cases with revision no more than 5 years after primary total hip replacement from January 2002 to June 2007. The reasons for revision were as follows: 2 cases were recurrent dislocation due to malposition of acetabular prosthesis; 5 cases were loosening of acetabular prosthesis; 1 case was abrasion of the native acetabulum by bipolar femoral head; 2 cases were periprosthetic femoral fractures and 1 case was periprosthetic infection. The average follow-up time was 36 months. Each patient was assessed according to Harris hip score. The revision procedures including liner only, acetabular prosthesis only, or both acetabular prosthesis and femoral prosthesis depending on the reasons for revision, two-stage revision was performed on 1 case with periprosthetic infection. RESULTS: The average of Harris hip score was increased from 46 (28 to 62) preoperatively to 86 (75 to 96) at follow up. The complication occurred in 2 cases: one was postoperative haematoma formation who was performed further surgery for clearance of haematoma, another was slight instability of the hip joint who was accepted skin traction for 3 weeks. CONCLUSIONS: The main reason for revision after primary total hip replacement is related to uncorrected insert of acetabular prosthesis. Improving surgical technique of insert of acetabular prosthesis is important in primary total hip replacement.


Assuntos
Artroplastia de Quadril , Complicações Pós-Operatórias , Falha de Prótese , Idoso , Feminino , Seguimentos , Humanos , Masculino , Pessoa de Meia-Idade , Complicações Pós-Operatórias/cirurgia , Reoperação , Estudos Retrospectivos , Resultado do Tratamento
11.
Zhongguo Gu Shang ; 21(3): 167-9, 2008 Mar.
Artigo em Chinês | MEDLINE | ID: mdl-19105427

RESUMO

OBJECTIVE: To evaluate therapeutic effects for reconstruction of anterior cruciate ligament(ACL)with hamstring tendon autografts and bioabsorbable interference screws fixation under arthroscopy. METHODS: Thirty-one patients with ACL rupture were verified through arthroscopy. There were 27 patients were male and 4 patients were female, ranging in age from 17 to 40 years,with an average of 25 years. Among the patients, 26 patients combined with meniscus injuries, 3 patients with injuries of articular cartilage and 16 patients with I to II degree degeneration of articular cartilage. All the patients were performed ACL reconstruction with hamstring tendon autografts under arthroscopy and the reconstructed ligaments were fixed with bioabsorbable interference screws. RESULTS: No severe complications occurred at early stage after operation. Thirty patients were followed up and ranged from 9 to 39 months,with an average of (19 +/- 9.0) months. Lysholm score significantly increased from average of 54.6 +/- 16.6 preoperatively to average of 92.5 +/- 5.7 at the end of follow-up period (t = 11.84, P < 0.01). Twenty-six patients restored to normal activity. CONCLUSION: ACL reconstructed with hamstring tendon autografts under arthroscopy has advantages of minimal trauma and satisfactory outcomes.


Assuntos
Ligamento Cruzado Anterior/cirurgia , Tendões/transplante , Adolescente , Adulto , Artroscopia , Feminino , Humanos , Masculino , Procedimentos de Cirurgia Plástica , Tendões/cirurgia , Transplante Autólogo , Resultado do Tratamento
12.
Life Sci ; 82(7-8): 393-401, 2008 Feb 13.
Artigo em Inglês | MEDLINE | ID: mdl-18191951

RESUMO

Interferonalpha (IFNalpha) induces cell cycle arrest and triggers apoptosis and chemosensitivity. But the mechanism of IFNalpha in regulating chemosensitivity has not been fully understood. To study whether IFNalpha affected chemosensitivity of osteosarcoma cells, we treated p53-wild U2OS cells and p53-mutant MG63 cells with IFNalpha and etoposide, alone or in combination, and then examined growth inhibition, cell cycle arrest and apoptosis. IFNalpha enhanced etoposide-induced growth inhibition and apoptosis in p53-wild U2OS cells but not p53-mutant MG63 cells in a dose- and time-dependent manner. Etoposide-induced G2/M phase arrest was also enhanced by IFNalpha. The enhanced apoptosis was associated with the accumulation of transcriptionally active p53 accompanied with increased Bax and Mdm2, as well as decreased Bcl-2. IFNalpha also activated caspases-3, -8 and -9 protein kinases and PARP cleavage in response to etoposide in U2OS cells. Moreover, the combination-induced cytotoxicity and PARP cleavage were significantly reduced by caspase pan inhibitor and p53 siRNA. Thus we conclude that IFNalpha enhances etoposide-induced apoptosis in human osteosarcoma U2OS cells by a p53-dependent and caspase-activation pathway. The proper combination of IFNalpha and conventional chemotherapeutic agents may be a rational strategy for the treatment of human osteosarcoma with functional p53.


Assuntos
Antineoplásicos Fitogênicos/farmacologia , Apoptose/efeitos dos fármacos , Neoplasias Ósseas/patologia , Etoposídeo/farmacologia , Interferon-alfa/farmacologia , Osteossarcoma/patologia , Proteína Supressora de Tumor p53/metabolismo , Neoplasias Ósseas/tratamento farmacológico , Neoplasias Ósseas/metabolismo , Linhagem Celular Tumoral , Sobrevivência Celular/efeitos dos fármacos , Combinação de Medicamentos , Humanos , Mutação , Osteossarcoma/tratamento farmacológico , Osteossarcoma/metabolismo , Proteínas Proto-Oncogênicas c-mdm2/metabolismo , Proteína Supressora de Tumor p53/genética , Proteína X Associada a bcl-2/metabolismo
13.
Zhonghua Wai Ke Za Zhi ; 45(18): 1279-83, 2007 Sep 15.
Artigo em Chinês | MEDLINE | ID: mdl-18067746

RESUMO

OBJECTIVES: To facilitate gene therapy research using recombinant adeno-associated virus type 2 (rAAV2) vector as gene transfer vehicle, and to construct a rAAV2 based vector carrying bone morphogenetic protein-7 (BMP7) and observe its expression in bone mesenchymal stem cells. METHODS: The coding sequence (1.3 kb) of BMP7 was amplified by polymerase chain reaction (PCR) from the pcDNA1.1(+) plasmid containing the human BMP-7 cDNA. After purified, the gene fragment was cloned into a plasmid pUC18 and termed plasmid pUC18-hBMP7. The recombinant pUC18-hBMP7 was digested by Kpn I and Sal I and further ligated to the pSNAV by T4DNA ligase. The resultant plasmid PSNAV-hBMP7 was transformed into DH5a Escherichia coli, and positive colonies were screened by PCR and digest with restriction enzyme to identify the correct recombinant clones. BHK-21 cells were transfected with the purified pSNAV-BMP7 plasmid according to a standard calcium phosphate precipitation method. The cells were then cultured in selection media containing 800 micro g/ml G418 (Gibco/BRL). G418-resistant BHK-21 cell clones were isolated and the integrity of hBMP7 gene was determined by PCR using the above PCR primers. To package the virus, stably transfected BHK-21 cells were subsequently infected with recombinant herpes simplex virus type 1 (rHSV-1). The collected cells were processed by chloroform treatment, PEG8000/NaCl precipitation and chloroform extraction for purification. The titer was determined using quantitative DNA dot blots and the purity was examined by sodium dodecyl sulphate polyacrylamide gel electrophoresis. Following infection with rAAV2-BMP7 at multiplicities of infection of 1 x 10(5) vector genomes per cell and subsequent culture, MSCs were assessed qualitatively for BMP7 production. RESULTS: Transient transfection showed an efficiency of 98.8% in MSCs. RT-PCR showed that MSCs had transcription of BMP7 that was enhanced by the gene transfer. BMP-7 expression in MSCs was identified by Western-blot. CONCLUSIONS: The hBMP7 recombinant adeno-associated virus vector is successfully constructed. The present in vitro study demonstrates that rAAV2-BMP7 could infect MSCs.


Assuntos
Proteínas Morfogenéticas Ósseas/genética , Dependovirus/genética , Vetores Genéticos/genética , Células-Tronco Mesenquimais/metabolismo , Fator de Crescimento Transformador beta/genética , Animais , Western Blotting , Células da Medula Óssea/citologia , Células da Medula Óssea/metabolismo , Proteína Morfogenética Óssea 7 , Proteínas Morfogenéticas Ósseas/metabolismo , Linhagem Celular , Células Cultivadas , Expressão Gênica , Células-Tronco Mesenquimais/citologia , Coelhos , Proteínas Recombinantes de Fusão/genética , Proteínas Recombinantes de Fusão/metabolismo , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Transfecção , Fator de Crescimento Transformador beta/metabolismo
14.
Acta Pharmacol Sin ; 28(11): 1835-41, 2007 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-17959036

RESUMO

AIM: To determine whether interferon-alpha(IFNalpha) can enhance doxorubicin sensitivity in osteosarcoma cells and its molecular mechanism. METHODS: Cell viability was evaluated using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay. Apoptosis was studied using Flow cytometry analysis, Hoechst33258 staining, DNA fragmentation assay, as well as the activation of caspase-3 and poly (ADP-ribose) polymerase. Protein expression was detected by Western blotting. The dependence of p53 was determined using p53-siRNA transfection. RESULTS: IFNalpha increased doxorubicin-induced cytotoxicity to a much greater degree through apoptosis in human osteosarcoma p53-wild U2OS cells, but not p53-mutant MG63 cells. IFNalpha markedly upregulated p53, Bax, Mdm2, and p21, downregulated Bcl-2, and activated caspase-3 and PARP cleavage in response to doxorubicin in U2OS cells. Moreover, the siRNA-mediated silencing of p53 significantly reduced the IFNalpha/doxorubicin combination-induced cytotoxicity and PARP cleavage. CONCLUSION: IFNalpha enhances the sensitivity of human osteosarcoma U2OS cells to doxorubicin by p53-dependent apoptosis. The proper combination with IFNalpha and conventional chemotherapeutic agents may be a rational strategy for improving the treatment of osteosarcoma with functional p53.


Assuntos
Antibióticos Antineoplásicos/farmacologia , Apoptose/efeitos dos fármacos , Doxorrubicina/farmacologia , Interferon-alfa/farmacologia , Osteossarcoma/tratamento farmacológico , Antibióticos Antineoplásicos/administração & dosagem , Linhagem Celular Tumoral , Sobrevivência Celular/efeitos dos fármacos , Sobrevivência Celular/genética , Doxorrubicina/administração & dosagem , Sinergismo Farmacológico , Genes p53/fisiologia , Humanos , Técnicas In Vitro , Interferon-alfa/administração & dosagem , Mutação , Osteossarcoma/metabolismo , Osteossarcoma/patologia , Ativação Transcricional
15.
Cancer Biol Ther ; 6(7): 1074-80, 2007 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-17611411

RESUMO

The tumor suppressor p14ARF, encoded by the INK4a/ARF locus, is often disrupted in human cancers, p14ARF triggers cell cycle arrest and sensitizes cells to apoptosis in the presence of collateral signals. To investigate the role of p14ARF in chemotherapeutic drugs-induced apoptosis, p14ARF was overexpressed by stable transfection in human osteosarcoma cell lines, U2OS (p53-wt/p14ARF-null) and MG63 (p53-mt/p14ARF-null). The results showed that ectopic p14ARF sensitized both cell lines to cisplatin-induced cytotoxicity and apoptosis. This sensitization of cisplatin-induced apoptosis was associated with upregulation of p53, Bax and p21 in U2OS cells. Conversely, such a result was not observed in MG63 cells. Moreover, the sensitization of cisplatin-induced cytotoxicity in U2OS cells was unaltered by p53 siRNA. Together, we show here p14ARF sensitizes human osteosarcoma cells to cisplatin-induced apoptosis in a p53-independent manner. Proper combinations of p14ARF gene transfer and conventional chemotherapy may be a valuable strategy in human osteosarcoma treatment.


Assuntos
Antineoplásicos/farmacologia , Apoptose/efeitos dos fármacos , Neoplasias Ósseas/tratamento farmacológico , Cisplatino/farmacologia , Osteossarcoma/tratamento farmacológico , Proteína Supressora de Tumor p14ARF/fisiologia , Proteína Supressora de Tumor p53/fisiologia , Neoplasias Ósseas/patologia , Linhagem Celular Tumoral , Humanos , Osteossarcoma/patologia , Proteínas Proto-Oncogênicas c-mdm2/análise , Transfecção , Proteína Supressora de Tumor p14ARF/genética , Proteína Supressora de Tumor p53/análise , Receptor fas/análise
16.
Acta Pharmacol Sin ; 28(6): 839-49, 2007 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-17506943

RESUMO

AIM: To determine whether adeno-associated virus (AAV)-2-mediated, bone morphogenetic protein (BMP)-7-expressing human adipose-derived mesenchymal stem cells (ADMS) cells would induce bone formation in vitro and in vivo. METHODS: ADMS cells were harvested from patients undergoing selective suction-assisted lipectomy and transduced with AAV carrying the human BMP-7 gene. Non-transduced cells and cells transduced with AAV serotype 2 carrying the enhanced green fluorescence protein gene served as controls. ADMS cells were qualitatively assessed for the production of BMP-7 and osteocalcin, and subjected to alkaline phosphatase (ALP) and Chinalizarin staining. A total of 2.5 x 10(6) cells mixed with type I collagen were implanted into the hind limb of severe combined immune-deficient (SCID) mice and subjected to a histological analysis 3 weeks post implantation. RESULTS: Transfection of the ADMS cells achieved an efficiency of 99% at d 7. Transduction with AAV2-BMP-7 induced the expression of BMP-7 until d 56, which was markedly increased by d 7. The cells were positively stained for ALP. Osteocalcin production and matrix mineralization further confirmed that these cells differentiated into osteoblasts and induced bone formation in vitro. A histological examination demonstrated that implantation of BMP-7-expressing ADMS cells could induce new bone formation in vivo. CONCLUSION: The present in vitro and in vivo study demonstrated that human ADMS cells would be a promising source of autologous mesenchymal stem cells for BMP gene therapy and tissue engineering.


Assuntos
Tecido Adiposo/metabolismo , Proteína Morfogenética Óssea 7/genética , Dependovirus/metabolismo , Terapia Genética/métodos , Células-Tronco Mesenquimais/fisiologia , Osteogênese/fisiologia , Animais , Proteína Morfogenética Óssea 7/metabolismo , Linhagem Celular , Forma Celular , Transplante de Células , Dependovirus/genética , Humanos , Imunofenotipagem , Masculino , Células-Tronco Mesenquimais/citologia , Camundongos , Camundongos SCID , Osteocalcina/metabolismo
17.
Acta Orthop ; 77(3): 512-8, 2006 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-16819694

RESUMO

BACKGROUND: As revision total knee arthroplasty surgery is becoming more common, it is necessary to evaluate how individual revision prosthesis systems perform in degenerative and inflammatory arthritides. In this study, results of the use of the Total Condylar III (TC III) system in osteoarthritis (55 knees) were compared to results of its use in inflammatory arthritis (16). METHODS: Patients were followed radiographically for 5.9 (3.0-10.2) years and clinically for 3.0 (0.2-6.8) years, using re-revision as the endpoint. RESULTS: At 1 year after revision and at final follow-up, the total Knee Society knee score, function score and range of motion had improved (p < 0.001) with no differences between osteoarthritis and inflammatory arthritis. No knee had definite component loosening, although 23 knees had asymptomatic radiolucent lines. Complications comprised 4 infections, 1 patellar pain syndrome and 1 rupture of the patellar tendon. Using any re-revision of the prosthesis as the endpoint, 5-year survival was 95% and 8-year survival was 94%. INTERPRETATION: Concentration of demanding revision knee arthroplasties to a few hands led to good or excellent knee joint knee score results in four-fifths of the patients, and showed good outcome with the TCIII system. In spite of ligamentous laxity, propensity to develop infections, bone destruction and poor general health, patients with inflammatory arthritis had results similar to those with osteoarthritis.


Assuntos
Artrite Reumatoide/cirurgia , Artroplastia do Joelho/efeitos adversos , Osteoartrite do Joelho/cirurgia , Adulto , Idoso , Artrite Reumatoide/diagnóstico por imagem , Feminino , Seguimentos , Humanos , Prótese do Joelho/efeitos adversos , Masculino , Pessoa de Meia-Idade , Osteoartrite do Joelho/diagnóstico por imagem , Falha de Prótese , Infecções Relacionadas à Prótese/microbiologia , Infecções Relacionadas à Prótese/cirurgia , Radiografia , Reoperação , Resultado do Tratamento
18.
J Bone Joint Surg Am ; 88(7): 1425-30, 2006 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-16818966

RESUMO

BACKGROUND: National and regional arthroplasty registries have been used to study the results of primary total knee arthroplasties. The purpose of this paper was to present the results of revision total knee replacements and describe predictors of survival of those replacements, with repeat revision as the end point. METHODS: The nationwide Finnish Arthroplasty Registry included 2637 revision total knee arthroplasties from 1990 through 2002. Survivorship of the revision total knee arthroplasties was analyzed, with repeat revision as the end point. The survivorship analyses comprised evaluations of the proportional hazards assumption followed by calculations of univariate and multivariate statistics and model diagnostics as appropriate. RESULTS: The survival rate following the revision total knee arthroplasties was 95% (95% confidence interval, 94% to 96%) at two years (1874 knees), 89% (95% confidence interval, 88% to 90%) at five years (944 knees), and 79% (95% confidence interval, 78% to 81%) at ten years (141 knees). Multivariate regression analysis showed the most significant predictors of prosthetic survival to be the age of the patient and the life in service of the primary total knee replacement (that is, the time between the primary total knee replacement and the revision). Survivorship was also significantly predicted by the year of the first revision total knee arthroplasty and the reason for the revision. CONCLUSIONS: An age greater than seventy years, revision five years or more after the primary arthroplasty, and absence of patellar subluxation are positive indicators of survival of a revision total knee replacement. We believe that normal aging as well as the deconditioning effect of disease (osteoarthritis and rheumatoid arthritis) and its treatment (primary total knee replacement) may lead to a reduced activity level, which, together with a presumed reluctance to operate on elderly patients, protects against repeat revisions. LEVEL OF EVIDENCE: Prognostic Level II. See Instructions to Authors for a complete description of levels of evidence.


Assuntos
Artroplastia do Joelho , Falha de Prótese , Sistema de Registros , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Feminino , Finlândia , Seguimentos , Humanos , Masculino , Pessoa de Meia-Idade , Desenho de Prótese , Reoperação , Fatores de Risco , Fatores de Tempo , Resultado do Tratamento
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