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1.
Chinese Journal of Microsurgery ; (6): 508-514, 2022.
Article in Chinese | WPRIM | ID: wpr-958395

ABSTRACT

Objective:To investigate the clinical efficacy of the vascularised semi-split fibular flap with lateral approach in ankle fusion.Methods:A total of 54 patients who underwent ankle fusion through the lateral menstrual approach by the Department of Trauma and Micro Orthopaedics, Zhongnan Hospital of Wuhan University from June 2015 to December 2020 was retrospectively analysed. Of the 54 patients, 27 patients who underwent ankle fusion with a semi-split fibular flap carrying blood supply were assigned to the fibular flap group, while other 27 patients who had ankle fusion with lateral plate were assigned to the plate group. The ankle fusions for both groups were performed by the same surgical team. The clinical efficacy of the 2 groups was evaluated by the success rate of bone fusion, the American Orthopedic Foot and Ankle Surgery(AOFAS) Ankle-Hindfoot Score System, and the Visual analog score(VAS). All the patients entered the postoperative follow-up at outpatient clinic.Results:The follow-up lasted for 12-24 months, with an average of 15 months. At 6 months after surgery, 24 patients in the fibular flap group achieved osseous union with 88.9% in the success rate of bone fusion. While 17 patients in the plate group achieved osseous union, with the success rate of bone fusion at 63.0%. There was a statistically significant difference between the groups( P<0.05). At 1 year after surgery, all 27 patients in the fibular flap group achieved bony union, with a 100% of the success rate of bone fusion, while 23 patients in the plate group achieved bony union at a 85.2% success rate of bone fusion, with 4 patients failed to heal. The difference was statistically significant( P<0.05). The AOFAS scores of the fibular flap group were 41.3±12.0, 65.6±5.6, and 79.1±7.0 before operation, at 6 months after operation and 1 year after operation, respectively, while the scores for the plate group at the same time were 40.8±11.3, 64.5±4.1 and 69.3±7.2. There was no significant difference in the scores between the groups before surgery and at 6 months after the operation ( P>0.05). The scores at 1 year after surgery in the fibular flap group was significantly higher than the plate group, and there was a statistically significant difference( P<0.05). The preoperative VAS score in the fibular flap group was 7.6±1.3, while it was 7.5±1.1 in the plate group. There was no significant difference between the groups in VAS score( P>0.05). However, the VAS score at 1 year after surgery was found at 1.8±0.9 in the plate group, and 1.9±0.8 in the fibular flap group and the difference between groups was statistically significant( P<0.05). Conclusion:The surgical procedure of semi-split fibular flap through the lateral approach has lower complications and higher success rate in ankle fusion in comparison with those of the fusion with lateral plate. Thus, further clinical investigations can be considered.

2.
Chinese Journal of General Practitioners ; (6): 743-747, 2022.
Article in Chinese | WPRIM | ID: wpr-957898

ABSTRACT

Clinical data of 14 patients with skin and soft tissue defects treated by progressive stretch skin suture (stretch suture) in Zhongnan Hospital of Wuhan University from September 2016 to January 2021 were retrospectively analyzed. There were 10 male and 4 female aged 13-73 years. The ankle and calf were the main defect sites with the defect area ranged from 2-3 cm×2-3 cm to 3-6 cm×5-9 cm. According to the wound skin condition, patients received progressive stretch suture 2-4 times after debridement and anti-infection treatment until the wounds were completely healed. Two patients with fractures underwent skin stretch after fracture fixation with external fixators. There were 4 patients with wound infection, including 2 cases infected with Staphylococcus aureus, 1 with Escherichia coli, and 1 with Enterobacter cloacae, all of whom received antibiotic therapy and infection was controlled and the progressive stretch suture was performed. In 3 patients with severe degloved skin injury, the infection was treated with debridement on the first day, and stretch suture was not performed until the second debridement. The soft tissue defect wound healed completely in 10-23 days, with an average of (17.2±3.8)days. None of the patients had skin necrosis during the stretch suture treatment. The postoperative follow-up time was more than 6 months and the skin softness, sensory function and blood supply of the patients were normal, and the average Vancouver Scar Scale score was 3.1, among whom 9 cases were scored as excellent (0-3 points) and 5 as good (4-7 points). The patients were satisfied with the effect of skin stretch,the study suggests that the progressive stretch skin suture technique is simple and effective in treatment of soft tissue defect.

3.
Chinese Journal of Orthopaedic Trauma ; (12): 503-509, 2022.
Article in Chinese | WPRIM | ID: wpr-956548

ABSTRACT

Objective:To access the efficacy of external fixation combined with Prontosan management for open fractures complicated with multidrug-resistant bacterial infection.Methods:A retrospective analysis was conducted of the data of 22 patients with open fracture complicated by multidrug-resistant bacterial infection who had been admitted to Department of Orthopedic Trauma and Microsurgery, Zhongnan Hospital of Wuhan University from January 2015 to January 2020. According to whether the Prontosan management was used or not, the patients were divided into 2 groups.The Protosan group [9 males and 3 females with an age of (44.6±13.1) years] were subjected to external fixation, vacuum sealing drainage(VSD) or conventional dressing changes, and at the same time Protosan management to flush the wound or Prontosan gel to change dressings.The control group[6 males and 4 females with an age of (45.1±11.7) years] were subjected to external fixation plus VSD or conventional dressing changes. Skin flaps or skin grafts were used to repair the wound immediately after the wound infection was controlled in both groups. The time for negative culture of the bacteria on the wound surface, number of debridement, survival of the skinflaps or grafts, and fracture union were recorded and compared between the 2 groups.Results:There was no significant difference in the preoperative general data between the 2 groups which were comparable( P>0.05).All patients were followed up for 10 to 24 months (mean, 14.5 months).In the Prontosan group, the number of debridement was 2.0 (2.0, 3.0) times, significantly fewer than that in the control group [4.0 (3.0, 4.0) times]; the time for negative bacterial culture (8.3±2.2) d, significantly shorter than that in the control group [(14.2±3.1) d]; the fracture union time (5.5±1.1) months, significantly shorter than that in the control group [(6.5 ±1.1) months]; the Samantha X-ray score at 6 months after operation 6.0 (5.0,6.0) points, significantly higher than that in the control group [5.2(4.5,5.5) points] (all P<0.05). Skin flaps or grafts survived in all the patients without any nonunion or chronic osteomyelitis. Conclusion:In the external fixation of open fractures complicated with multidrug-resistant bacterial infection, combination with Prontosan management can effectively control infection and promote granulation, early wound healing and fracture union.

4.
Chinese Journal of Medical Instrumentation ; (6): 471-475, 2020.
Article in Chinese | WPRIM | ID: wpr-880393

ABSTRACT

A clinical information navigation system based on 3D human body model is designed. The system extracts the key information of diagnosis and treatment of patients by searching the historical medical records, and stores the focus information in a predefined structured patient instance. In addition, the rule mapping is established between the patient instance and the three-dimensional human body model, the focus information is visualized on the three-dimensional human body model, and the trend curve can be drawn according to the change of the focus, meanwhile, the key diagnosis and treatment information and the original report reference function are provided. The system can support the analysis, storage and visualization of various types of reports, improve the efficiency of doctors' retrieval of patient information, and reduce the treatment time.


Subject(s)
Humans , Diagnosis, Computer-Assisted , Medical Informatics Applications , Models, Anatomic , Software
5.
West China Journal of Stomatology ; (6): 174-179, 2019.
Article in Chinese | WPRIM | ID: wpr-772679

ABSTRACT

OBJECTIVE@#To evaluate and analyze the accuracy of three-dimensional camera system acquisition based on stereophotography for the photographic acquisition of images of facial deformities.@*METHODS@#3D digital models of 45 waxen facial models of patients with facial deformities were obtained via a 3D camera system based on stereophotography. A total of 19 feature lines were measured on each 3D model by using the software. The measurements taken by the coordinate-measuring machine were accepted as standard values. Statistical analysis was performed to evaluate the accuracy of the system and the corresponding factors.@*RESULTS@#Statistical analysis results showed that the measured values of the characteristic distance obtained by 3D camera system were statistically different from the standard values (P0.05). The facial regions significantly influenced measuring error (P0.05). The middle facial regions had significant difference with bilateral facial regions on the percentage of measurement values (P0.05).@*CONCLUSIONS@#The accuracy of 3D camera system based on stereophotography for the photographic acquisition of facial deformities are influenced by the morphology of the facial deformities and facial regions. Moreover, the measuring error is acceptable in clinical settings.


Subject(s)
Humans , Craniofacial Abnormalities , Diagnostic Imaging , Imaging, Three-Dimensional , Reproducibility of Results , Software
6.
Chinese Journal of Orthopaedic Trauma ; (12): 581-585, 2019.
Article in Chinese | WPRIM | ID: wpr-754766

ABSTRACT

Objective To investigate the surgical outcomes of one-stage repair of the ankle fracture combined with grade Ⅲ injury to the lateral ligament.Methods From April 2015 to December 2017,23 patients with ankle fracture and lateral ligament injury were treated at Department of Traumatic Reconstruction Surgery/Foot and Ankle Surgery,Zhongnan Hospital.There were 15 males and 8 females,aged from 18 to 65 years (average,37.8 years).After intraoperative treatment of their fractures,22 ruptures of the ligamentous insertion were repaired with suture anchor and one rupture of the middle ligament with Internal Brace.Routine postoperative X-rays,physical examination and Sefton grading system were used to evaluate the stability of the ankle joint and the subtalar joint.The Karlsson-Peterson and American Orthopedic Foot and Ankle Society (AOFAS) scores were used to evaluate the function and pain of the ankle.Results The follow-up for this cohort ranged from 12 to 24 months (average,13.9 months).The bone union time ranged from 6 to 24 weeks,averaging 10.1 weeks.There was no chronic instability of lateral ankle or subtalar joint in all the cases.Two cases developed osteochondral lesion of the medial talus which was treated with micro-fracture surgery one year after injury;one case suffered subtalar arthritis which was treated by subtalar arthrodesis 14 months after injury.Their Karlsson-Peterson scores averaged 84.6 points and AOFAS ankle-hindfoot scores 93.7 points with 19 excellent and 4 good cases.The Sefton grading system gave 9 cases of grade Ⅰ and 14 ones of grade Ⅱ.Conclusion One-stage repair of the ankle fracture combined with grade Ⅲ injury to the lateral ligament can lead to the stability of the ankle joint and the subtalar joint and improved function of the ankle.

7.
Chinese Journal of Trauma ; (12): 1007-1013, 2018.
Article in Chinese | WPRIM | ID: wpr-707396

ABSTRACT

Objective To investigate the clinical efficacy of three-stage Masquelet technique in the treatment of infective bone defects of foot and ankle.Methods A retrospective case series study was conducted on the clinical data of 19 patients with infective bone defects of foot and ankle admitted to Zhongnan Hospital of Wuhan University from December 2014 to October 2017.There were 15 males and four females,aged 18-68 years [(39.6 ± 12.3)years].Among the patients,16 patients were infected with bacteria and three patients were infected with Mycobacterium tuberculosis.The infection involved humeral end,talus and surrounding joints in 14 patients,internal hemorrhoids in two patients,midfoot and interphalangeal joints in one patient,and humerus and metatarsophalangeal joints in two.The operation included three stages:the first stage was thorough debridement,supplemented with negative pressure closed drainage (VSD) and continuous washing to clarify the pathogenic bacteria;the second stage was to fill the bone defect with targeted antibiotic bone cement to prevent or treat infection;in the third stage,after filling the antibiotic bone cement for 3 months with no sign of local wound infection,the bone cement was taken out,and the bone reconstruction operation was performed by means of internal fixation and bone grafting.The flap survival and wound healing were observed,and the time of fracture healing was recorded.The American Orthopaedic Foot and Ankle Society (AOFAS) score and the Visual Analogue Score (VAS) were used to evaluate the improvement of the function of the foot and ankle before operation and at the last follow-up,and the bone healing was evaluated according to the radiographic union scale in tibial (RUST) fractures.Results The patients were followed up for 9-12 months [(11.1 ±1.0) months].Two patients with soft tissue defects caused by preoperative infection and necrosis received posterior tibial artery perforator flap and anterolateral thigh flap repair in the second stage,and the flaps all survived.The postoperative bone healing time was 3 7 months [(3.5 ± 1.4)months].Nineteen patients underwent three-stage surgery,and the ankle and foot wound or sinus tract were all healed,with no infection recurrence during follow-up.At the last follow-up,the AOFAS score was improved significantly from preoperative (36.3 s-12.1) points to (71.4 ± 5.7) points (P < 0.05).The VAS was decreased significantly from preoperative (5.3 ± 1.2) points to (1.4 ± 0.9) points (P < 0.05).The RUST bone healing score at the last follow-up was 8-12 points [(10.2 ± 1.1) points].Conclusion In treating the infective bone defects of foot and ankle,the three-stage Masquelet technique can effectively control infection,facilitate wound healing,promote bone union,and improve foot and ankle function.

8.
Chinese Journal of Health Policy ; (12): 40-46, 2018.
Article in Chinese | WPRIM | ID: wpr-703598

ABSTRACT

This paper systematically reviews the latest and relevant literatures and policy documents on the in-tegrated health services in Canada in recent years. Therefore, it summarizes the practice and mode of integrated serv-ice delivery in Ontario, Alberta and Quebec wherein the integration among health organization, health service team, and a series of health services are included. The contributing factors and impeding factors ( the barriers) of organiza-tional integration and specific integration strategy were summarized. Finally, according to the actual practical situa-tion, it is proposed that China should adhere to the government-led approach in promoting the integration of health services, and give a full play to the positive role of the market mechanism. Through strengthening the network man-agement and group service of primary health services, emphasis will be put on health services of population groups and specific diseases. Therefore, integration will be regarded as a strategic priority, increasing incentives and boos-ting promotion of nursing personnel on the process of Integrated Service Delivery, building the health information sys-tem that is conducive to integration in order to continuously advance Hierarchical Diagnosis and bridge the fragmented service system. This will help in providing residents with personalized, convenient, comprehensive, and continuous health services.

9.
Chinese Journal of Microbiology and Immunology ; (12): 456-460, 2018.
Article in Chinese | WPRIM | ID: wpr-711428

ABSTRACT

Objective To investigate the influence on anti-HCV antibody levels in spontaneous HCV seroconverters co-infected with HIV. Methods A retrospective study was conducted on people with a history of blood donation in Wangying Village,Shangcai County,Henan Province in 2009 and 2017. Accord-ing to the infection status in 2009,patients who were positive for anti-HCV antibody were divided into four groups:HIV-negative chronic HCV infection group (HCVc),HIV-negative spontaneous HCV clearance group (HCVr),HIV-positive chronic HCV infection group (HIV+HCVc),HIV-positive spontaneous HCV clear-ance group ( HIV+HCVr). All patients were followed up in 2017 and those who were lost to follow-up, received HCV treatment or were reinfected with HCV (only for those of HCV seroconverters) were excluded from this study. Altogether 167 patients met the inclusion criteria (HCVc:n=65;HCVr:n=34;HIV+HCVc:n=44;HIV+HCVr:n=24). A horizontal comparison of anti-HCV antibody levels among the above four groups in 2009 and a longitudinal comparison of changes in anti-HCV antibody in each group from 2009 to 2017 were respectively conducted. Results The horizontal comparison indicated that the levels of anti-HCV antibody were higher in chronic HCV-infected patients than in HCV seroconverters no matter whether they were co-infected with HIV or not (both P<0. 000 1). After comparison of anti-HCV antibody titers in 2017 and 2009,no significant changes were found in HCVc or HIV+HCVc group. The levels of anti-HCV antibody in HCVr and HIV+HCVr groups decreased significantly from 2009 to 2017 ( both P<0. 000 1). HIV+HCVr group showed a faster decline in anti-HCV antibody level than HCVr group (P=0. 003 9). Significant nega-tive correlations between the decline speed in anti-HCV antibody sample/cut-off ( S/CO) values and the initial anti-HCV antibody S/CO values (in 2009) were found in both HCVr (r=-0. 517 7, P=0. 001 7) and HIV+HCVr groups (r=-0. 753 2, P<0. 000 1). The decline speed in anti-HCV antibody in HIV+HCVr patients was found to be negatively correlated with their CD4+T cell counts in 2009 ( r=-0. 563 8, P=0. 004 1). Moreover,the seroreversion rate of anti-HCV antibody in patients of the HIV+HCVr group was higher than that of HCVr group (P=0. 027 5). Conclusion HIV co-infection can accelerate the decline of anti-HCV antibody in spontaneous HCV seroconverters. This study indicates that in a large-scale retrospective epidemiological investigation especially for HIV-infected populations, the prevalence of anti-HCV antibody may be underestimated.

10.
Chinese Journal of Microbiology and Immunology ; (12): 1-6, 2018.
Article in Chinese | WPRIM | ID: wpr-711359

ABSTRACT

Objective To analyze the possibility of using intracellular cytokine staining (ICS) to evaluate NK cell-mediated antibody-dependent cellular cytotoxicity (ADCC) and to detect the changes in ADCC activity among patients with chronic HIV and/or HCV infection. Methods Flow cytometry was per-formed to determine the percentages of NK cells and the expression of NK cell receptors. ImageStreamX MarkⅡ system was used to identify the expression of CD3, CD56, CD16 and CD32 on CD56brightNK and CD56dimNK subsets. Degranulation process and cytokine production in NK cells were detected using an anti-gen-antibody complex model of P815/Ab in combination with ICS. Differences in NK cell-mediated ADCC were evaluated among patients infected with HIV and/or HIV and healthy subjects by flow cytometry. Re-sults The percentages of CD107a+and IFN-γ+NK cells were positively correlated with the decrease of mean fluorescence intensity (MFI) of CD16. ICS assay revealed a positive correlation between the secretion of CD107a and IFN-γ by NK cells. CD16 was highly expressed in CD56dimNK cells. The ADCC mediated by CD56dimNK cells was stronger than that mediated by CD56brightNK cells. The rate of target cell lysis detected by rapid fluorescence assay was positively correlated with the percentage of CD107a+/IFN-γ+NK cells meas-ured by ICS. NK cell-mediated ADCC was suppressed in patients with chronic HIV and/or HCV infection. Conclusion This study suggests that ICS assay could be used to evaluate NK cell-mediated ADCC. It also reveals that NK cell-mediated ADCC is suppressed in patients with chronic HIV and/or HCV infection.

11.
Chinese Journal of Microsurgery ; (6): 538-543, 2018.
Article in Chinese | WPRIM | ID: wpr-735006

ABSTRACT

Objective To investigate the clinical effectivity of the muscular flap transposition and induced membrane technique in the emergency treatment for the limb salvage of Gustilo type Ⅲ B/C open fracture of lower leg. Methods From July, 2015 to December, 2017, 10 cases of Gustilo type Ⅲ B/C fracture of lower leg with bone defects were performed limb salvage surgery. Induced membrane technique was used to fill the bone defects in the emergency room.The gastrocnemius and/or soleus muscular flaps were transposed to cover the bone cement or ex-posed bone simultaneously in emergence treatment. After the wound healed completely, traditional bone grafting was used to repair the bone defects. There were 4 cases of Gustilo type Ⅲ B and 6 cases of Gustilo type Ⅲ C. The aver-age length of bone defect was (5.25±1.70) cm ranging from 3.0 cm to 11.0 cm. The gastrocnemius medial head flaps were performed in 5 cases, the combined application with the gastrocnemius medial head flaps and the medial hemimuscular flaps of soleus were performed in 2 cases, and medial hemimuscular flaps of soleus were transposed in 3 cases. Results The wounds in 6 cases were healed at one stage, but 2 cases healed by dressing because the exudate after skin grafting.In 1 case, the cross-leg flap was used to cover the exposed bone cement due to the necro-sis of soleus flap. The other 1 was performed the transposition of the lateral gastrocnemius flaps because the exposure of bone cement after the necrosis of the upper and lateral muscles in lower leg. In the second-stage, the bone defects were reconstructed by traditional bone grafting. The average healed time of bone was 7.2 months ranging from 5 months to 9 months. At the last followed-up time, all patients recovered their function of weight-bearing. The Paley's score of the adjacent joints: excellent in 8 cases and good in 2 cases. Conclusion The combination with induced membrane technique and local muscular flap transposition in emergency surgery is an effective method to limb salvage for the Gustilo type Ⅲ B/C open fracture of lower leg.

12.
Chinese Journal of Microbiology and Immunology ; (12): 305-309, 2016.
Article in Chinese | WPRIM | ID: wpr-486725

ABSTRACT

Objective To establish a TaqMan probe-based real-time fluorescent quantitative PCR ( real-time PCR) for the quantitative and rapid detection of viral reservoir in peripheral blood mononuclear cells (PBMCs) isolated from rhesus monkeys with simian immunodeficiency virus (SIV) infection and to evaluate its preliminary application. Methods A pair of primers and one TaqMan probe were designed ac-cording to the conserved sequence of SIVmac239 strain for real-time PCR amplification. A length of 2 090 bp of nucleotide fragment was digested from the plasmid p239SpSp5 containing 5′-end long segments of SIV-mac239 strain by restriction enzymes EcoRⅠand SpeⅠ. The standards used for quantitative detection of SIV DNA in peripheral blood samples were prepared by a 10-fold serial dilution and used for graphing the stand-ard curve. The numbers of SIV DNA ( copies per 106 PBMCs) in rhesus monkeys during acute and chronic phases of SIVmac239 infection were determined and the virological characteristics of SIV DNA at different phages of infection were analyzed. Results A linear positive correlation between cycle threshold ( Ct) val-ues and concentrations (10 copies/μl to 109 copies/μl) of the standards was found. High levels of SIV DNA were monitored in SIV-infected monkeys 14 to 22 days after acute infection. The levels of SIV DNA in the acute phase of infection were about 1 to 2 logs higher than those in the chronic phase of infection. The num-bers of SIV DNA ( copies per 106 PBMCs) were 1 log lower than the SIV viral load in peripheral blood of the same monkey. The ratios of SIV DNA load to SIV RNA load ( DNA/RNA) in chronic phase of infection were higher than those in the acute phase. Conclusion The established TaqMan probe-based real-time fluorescent quantitative PCR was a highly sensitive and specific assay for the detection of SIV DNA with an advantage of wide linear range. It could be used for the quantitative evaluation of latent reservoirs of SIV.

13.
Journal of Zhejiang University. Science. B ; (12): 923-930, 2008.
Article in English | WPRIM | ID: wpr-359340

ABSTRACT

<p><b>BACKGROUND</b>A major shortcoming in tissue engineered blood vessels (TEBVs) is the lack of healthy and easily attainable smooth muscle cells (SMCs). Smooth muscle progenitor cells (SPCs), especially from peripheral blood, may offer an alternative cell source for tissue engineering involving a less invasive harvesting technique.</p><p><b>METHODS</b>SPCs were isolated from 5-ml fresh rat peripheral blood by density-gradient centrifugation and cultured for 3 weeks in endothelial growth medium-2-MV (EGM-2-MV) medium containing platelet-derived growth factor-BB (PDGF BB). Before seeded on the synthesized scaffold, SPC-derived smooth muscle outgrowth cell (SOC) phenotypes were assessed by immuno-fluorescent staining, Western blot analysis, and reverse transcription polymerase chain reaction (RT-PCR). The cells were seeded onto the silk fibroin-modified poly(3-hydroxybutyrate-co-3-hydroxyhexanoate) (SF-PHBHHx) scaffolds by 6x10(4) cells/cm2 and cultured under the static condition for 3 weeks. The growth and proliferation of the seeded cells on the scaffold were analyzed by 3-(4,5-dimethylthiazol-2-yl)-diphenyltetrazolium bromide (MTT) assay, scanning electron microscope (SEM), and 4,6-diamidino-2-phenylindole (DAPI) staining.</p><p><b>RESULTS</b>SOCs displayed specific "hill and valley" morphology, expressed the specific markers of the SMC lineage: smooth muscle (SM) alpha-actin, calponin and smooth muscle myosin heavy chain (SM MHC) at protein and messenger ribonucleic acid (mRNA) levels. RT-PCR results demonstrate that SOCs also expressed smooth muscle protein 22alpha (SM22alpha), a contractile protein, and extracellular matrix components elastin and matrix Gla protein (MGP), as well as vascular endothelial growth factor (VEGF). After seeded on the SF-PHBHHx scaffold, the cells showed excellent metabolic activity and proliferation.</p><p><b>CONCLUSION</b>SPCs isolated from peripheral blood can be differentiated into the SMCs in vitro and have an impressive growth potential in the biodegradable synthesized scaffold. Thus, SPCs may be a promising cell source for constructing TEBVs.</p>


Subject(s)
Animals , Rats , 3-Hydroxybutyric Acid , Chemistry , Blood Vessels , Cell Biology , Caproates , Chemistry , Cell Adhesion , Cell Differentiation , Cell Proliferation , Immunophenotyping , Microscopy, Electron, Scanning , Muscle, Smooth, Vascular , Cell Biology , Myocytes, Smooth Muscle , Cell Biology , RNA, Messenger , Tissue Engineering , Vascular Endothelial Growth Factor A , Genetics
14.
Chinese Medical Journal ; (24): 696-702, 2007.
Article in English | WPRIM | ID: wpr-344827

ABSTRACT

<p><b>BACKGROUND</b>Tissue-engineered heart valves have the potential to overcome the limitations of present heart valve replacements. This study was designed to develop a tissue engineering heart valve by using human umbilical cord blood-derived endothelial progenitor cells (EPCs) and decellularized valve scaffolds.</p><p><b>METHODS</b>Decellularized valve scaffolds were prepared from fresh porcine heart valves. EPCs were isolated from fresh human umbilical cord blood by density gradient centrifugation, cultured for 3 weeks in EGM-2-MV medium, by which time the resultant cell population became endothelial in nature, as assessed by immunofluorescent staining. EPC-derived endothelial cells were seeded onto the decellularized scaffold at 3 x 10(6) cells/cm(2) and cultured under static conditions for 7 days. Proliferation of the seeded cells on the scaffolds was detected using the MTT assay. Tissue-engineered heart valves were analyzed by HE staining, immunofluorescent staining and scanning electron microscopy. The anti-thrombogenic function of the endothelium on the engineered heart valves was evaluated by platelet adhesion experiments and reverse transcription-polymerase chain reaction (RT-PCR) analysis for the expression of endothelial nitric oxide synthase (eNOS) and tissue-type plasminogen activator (t-PA).</p><p><b>RESULTS</b>EPC-derived endothelial cells showed a histolytic cobblestone morphology, expressed specific markers of the endothelial cell lineage including von Willebrand factor (vWF) and CD31, bound a human endothelial cell-specific lectin, Ulex Europaeus agglutinin-1 (UEA-1), and took up Dil-labeled low density lipoprotein (Dil-Ac-LDL). After seeding on the decellularized scaffold, the cells showed excellent metabolic activity and proliferation. The cells formed confluent endothelial monolayers atop the decellularized matrix, as assessed by HE staining and immunostaining for vWF and CD31. Scanning electron microscopy demonstrated the occurrence of tight junctions between cells forming the confluent monolayer. Platelets adhesion experiments suggested that the neo-endothelium was non-thrombogenic. The expression levels of eNOS and t-PA genes in the neo-endothelium were quite similar to those in human umbilical vein endothelial cells.</p><p><b>CONCLUSIONS</b>EPCs isolated from the human umbilical cord blood can differentiate into endothelial cells in vitro and form a functional endothelium atop decellularized heart valve scaffolds. Thus, EPCs may be a promising cell source for constructing tissue-engineered heart valves.</p>


Subject(s)
Animals , Humans , Cell Proliferation , Endothelial Cells , Cell Biology , Metabolism , Heart Valve Prosthesis , Heart Valves , Cell Biology , Metabolism , Immunohistochemistry , Microscopy, Electron, Scanning , Nitric Oxide Synthase Type III , Genetics , Metabolism , Platelet Aggregation , Reverse Transcriptase Polymerase Chain Reaction , Stem Cells , Cell Biology , Metabolism , Swine , Tissue Engineering , Methods , Tissue Plasminogen Activator , Genetics , Metabolism , Umbilical Cord , Cell Biology
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