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23 July 2020, Volume 10 Issue 6 Previous Issue    Next Issue

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Removal of Ofteratoma-forming Cells from Differentiated Embryonic Stem Cells Based on PECAM-1 Expression WANG Ling,TANG Zhengya,FU wei,ZHANG Wenjie 2014, 10 (6):  301-304.  DOI: 10.3969/j.issn.1673-0364.2014.06.02 Abstract ( 325 )   Objective To prevent teratoma formation by removal of the residual undifferentiated cells from differentiated mouse embryonic stem cells （mESCs） based on PECAM-1 expression, to hopefully promote the safe use of ESCs-based treatment in future. Methods Mouse R1 ESCs were cultured in suspension to form embryoid bodies （EBs） in the absence of leukemia inhibitory factor for 6 days. EBs were then digested into single cells and sorted by magnetic activated cell sorting （MACS） based on PECAM-1 expression. Total of 2×10^6 PECAM-1^＋ and PECAM-1^- cells were injected subcutaneously into nude mice respectively. Teratoma formation was measured after 6-8 weeks. Results Seven out of 8 injected points formed teratoma in the PECAM-1^＋ cells after 8 weeks of injection, with a tumor formation rate of 87.5%. While only 1 out of 8 injected points formed teratoma in the PECAM-1^- cells, with a tumor formation rate of 12.5%. A significantly difference was observed between PECAM-1 positive and negative cells （P=0.01）. Conclusion PECAM-1 is a specific marker for residual cells, which could be utilized to remove residual undifferentiated ESCs from in vitro differentiated ESCs, and eventually solve the tumorigenicity problem of ESCs. Related Articles | Metrics

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Low-Molecule-Weight Hydrogel M2 and Cross-Lincked Hyaluronic Acid as Injectable Cartilage Regeneration Scaffolds YIN Zongqi,LI Ping,LI Dan,LIU Yi,WANG Zhenxing,LIU Yu,FENG Chuanliang,ZHOU Guangdong 2014, 10 (6):  305-313.  DOI: 10.3969/j.issn.1673-0364.2014.06.002 Abstract ( 296 )   Objective To explore the feasibility of low-molecule-weight hydrogel M2 and cross-lincked hyaluronic acid （HA） as injectable materials for cartilage tissue engineering in nude mice. Methods The auricular chondrocytes of newborn pig were isolated and expanded. The cells were mixed with M2 hydrogel in 50×10^6 cells/mL and 100×10^6 cells/mL, and were mixed with HA in 100 ×10^6 cells/mL, followed by subcutaneous injection into nude mice. M2 hydrogel and HA were also injected as control groups. After 8 weeks, the samples were harvested and evaluated for cartilage formation by gross view, histochemical examination, wet weight test, mechanical analysis and glycosaminoglycan （GAG） quantification. Results All three groups formed cartilage-like tissues. Samples in M2 group with high cell density （100×10^6 cells/mL） were the largest in size with stiffest texture, smoothest surface, and most abundant lacuna structures. Samples in M2 group with low cell density （50 ×10^6 cells/mL） also formed cartilage-like tissue with stiff texture, but their size shrank obviously; HA group, however, formed immature cartilaginous tissue with poor rigidity, cellularization and cartilage specific matrix expression. The corresponding quantitative data further demonstrated that samples in M2 group with high cell density had the highest GAG content and strongest mechanical property. Statistically significant differences were observed among 3 groups. No cartilage-like tissue were formed in the 2 control groups. Conclusion Low-molecule-weight hydrogel M2 has better biocompatibility than hyaluronic acid, and M2 can be more suitable for TE cartilage construction. Related Articles | Metrics

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Human Adipose-De rived Stem Cells and its Biocompatibility with Bladder Acellular Matrix Graft-Silk Fibroin Bilayer Scaffold ZHAO Yang,WU Jiasheng,ZHOU Zhe,ZHOU Juan,ZHANG Ming,LI Wei,WANG Zhong,SUN Kang,LU Mujun 2014, 10 (6):  309-313.  DOI: 10.3969/j.issn.1673-0364.2014.06.003 Abstract ( 308 )   Objective To observe the growth of human adipose-derived stem cells （hASCs） in bladder acellular matrix graft-silk fibroin （BAMG-SF） bilayer scaffold and to analyze the biological compatibility of BAMG-SF with hASCs. Methods hASCs were isolated from human subcutaneous adipose tissue after collagenase digesting, filtrating and centrifuging, then cultured in the leaching solution of BAMG-SF. The cytotoxicity of scaffold was evaluated by CCK-8 cell viability assay, and the growth curves were also observed. Surface morphology on BAMG-SF was observed by scanning electron microscopy （SEM）. The hASCs of passage 3 were seeded onto the BAMG-SF bilayer scaffolds for 1 week, then the BAMG-SF bilayer scaffolds seeded with hASCs were transplanted into nude mouse for 1 week or 2 weeks. The growth of cells in BAMG-SF biomaterials was observed by HE staining. The species origin of these cells in the BAMG-SF scaffolds cultured in vivo was detected by Immunofluorescence. Results hASCs maintained high proliferation rate in the leaching solution of BAMG-SF and the BAMG-SF scaffolds were nontoxic absolutely. According to the growth curves of hASCs cultured in the leaching solution of the BAMG-SF and DMEM, BAMG-SF scaffolds were conducive to the growth of hASCs. The histological study found that hASCs could grow into the space of the BAMG-SF scaffolds after cultured in vitro and in vivo. There were more cells in the scaffolds cultured in vivo than in vitro. Immuno-fluorescence suggested that some of the cells inside the scaffolds were hASCs. Conclusion BAMG-SF bilayer scaffolds are nontoxic and have a good biocompatibility with hASCs, which can be used as a vehicle for hASCs in bladder defect reconstruction. Related Articles | Metrics

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Effectiveness of Umbilical Cord Mesenchymal Stem Cells in Treating Multiple System Atrophy-Cerebellar Type LIU Jing,HAN Dongmei,DING Li,XUE Mei,WANG Zhidong,YAN Hongmin,ZHU Ling,ZHENG Xiaoli,DONG Lei,GUO Zikuan,WANG Hengxiang 2014, 10 (6):  314-317.  DOI: 10.3969/j.issn.1673-0364.2014.06.004 Abstract ( 692 )   Objective To observe the effectiveness of intrathecal injection with umbilical cord mesenchymal stem cells （UC-MSCs） in treating multiple system atrophy-cerebellar type （MSA-C）. Methods From December 2009 to April 2012, 40 patients with MSA-C were given UC-MSCs intrathecal injection with the dosage of 1 ×10^6 cells/Kg for once a week, 4 times as a course. Twenty-nine patients were treated with 1 course, 9 patients with 2 courses and 2 patients with 3 courses. International Cooperative Ataxia Rating Scale （ICARS） and Activity of Daily Living Scale （ADL） were used to evaluate patients＇ neural function and ability of daily living. Patients were followed up after treatment. Results In 53 courses, 47 courses are effective （the effective rate was 88.68%）. Compared with the ICARS and ADL scores before treatment, patients’ scores were significantly decreased 1 month after treatment （P〈0.01）. In most effective cases, clinical symptoms such as unstable walking and standing, slow movement, upper limb fine motor disorder, writing difficulties, dysarthria, eye movement disorders were improved. During treatment, common adverse effects were observed, as dizziness （4 cases）, low back pain （1 cases）, headache （2 case） and fever （2 cases）, all these symptoms disappeared within 1-3 days. No treatment-related adverse events were observed during 27-55 months （median 40 months） of follow-up. The stable effective period of patients ws 2-13 months （average 5.88±2.86 months）, then progressed. Conclusion UC-MSCs intrathecal injection is safe, and can ameliorate clinical symptoms to some extent. Multiple courses of treatment can further improve neurological function and delay illness progression in most patients. Related Articles | Metrics

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Effect and M echanism of TGF-β Mediated Signal Pathway Modulated by microRNA-21 in Hypertrophic Scars ZHANG Qi,WANG Chen 2014, 10 (6):  318-323.  DOI: 10.3969/j.issn.1673-0364.2014.06.005 Abstract ( 308 )   Objective To evaluate the effect and mechanism of microRNA-21 in hypertrophic scars （HS） formation and to provide novel target in biological control of HS. Methods Normal skin tissue and HS tissue were harvested. Collagen quantitative detection and histological observation were carried out. The expression of collagen 1A1 （Col1A1）, collagen 3A1 （Col3A1）, Fibronectin （FN） and α-SMA were detected by quantitative real-time PCR. The expression of microRNA-21 mRNA in normal skin tissue and HS tissue were also detected. Cells isolated from normal skin tissues and HS tissues were cultured, the microRNA-21 antisense RNA interference vector were constructed, while TGF-β was added at the same time. Furthermore, the effects of inhibitor-microRNA-21 and TGF-β on the biological characteristics of fibroblasts were observed. Results The content of collagen in hypertrophic scar was higher than that in normal skin. The expression of extracellular matrix such as Col1A1, Col3A1, FN and α-SMA in scar tissue were significantly increased. The expression of microRNA-21 in HS tissue was higher than in normal tissue. TGF-βenhanced the expression of microRNA-21 in fibroblasts of HS tissues, while the inhibition of microRNA-21 decreased the expression of those scar-related genes. Conclusion The skin after wound healing could increase the expression of microRNA-21 which promotes the synthesis of extracellular matrix, and TGF-βmay further strengthen the role of microRNA-21 in the formation of HS. Related Articles | Metrics

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Chondrogenic Differentiation of Hypertrophic Scar -Derived Fibroblasts Initiated by Cartilage -derived Morphogenetic Protein 1 in Vivo SHEN Congcong,CHAI Gang,QU Miao,HOU Yikang,XU Yourong,ZHANG Yan 2014, 10 (6):  324-328.  DOI: 10.3969/j.issn.1673-0364.2014.06.006 Abstract ( 339 )   Objective To explore the chondrogenesis potential of hypertrophic scar fibroblasts （HSFBs） induced by cartilage-derived morphogenetic protein 1 （CDMP1） in vivo. Methods Hypertrophic scar fibroblasts （HSFBs） were isolated from discarded hypertrophic scar. HSFBs combining with PGA/PLA scaffold as HSFBs- PGA/PLA complex were induced by CDMP1 （100 ng/mL） for 2 weeks. Then the complex were transplanted into nude mice as induced group （n=10）. The complex that not induced were treated as negative control, named as un-induced group （ n=4）, while the chondrocytes-PGA/PLA complex were treated as positive control, named as chondrocyte group （n=4）. After 4 and 8 weeks, the complex in each group was harvested for wet weight test, proteoglycan （GAG） content test, HE staining, Safranine-O staining and immunochemistry staining. Results After 4 and 8 weeks in vivo, the wet weight and GAG content in induced group were both higher than in un-induced group （P〈0.05）. In induced group, after cultured for 4 weeks in vivo, chondrocyte-like lacuna structure was observed by HE staining, uniform distribution of GAG was observed by Safranine-O staining, and positive expression of collagen type Ⅱ was revealed by immunohistochemical staining. After 8 weeks of culturing in vivo, chondrocyte-like structure in induced group was more mature and more in line with the structure distribution of cartilage. Conclusion HSFBs combining with PGA/PLA had the potential to develop into polygonal chondrocyte-like tissue by the induction of CDMP 1. Related Articles | Metrics

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Closure of Large Defects after Microcystic Lymphatic Malformations Using Lateral Intercostal Artery Perforator Flap WANG Haizhen,CUI Chunxiao,JIANG Zhaohua,LI Shengli,CAO Weigang 2014, 10 (6):  329-338.  DOI: 10.3969/j.issn.1673-0364.2014.06.007 Abstract ( 312 )   Objective To evaluate the feasibility of complete removal of the multifocal and extensive lesions by accurate imaging diagnosis and to repair large skin defects due to complete resection of microcystic LMs using lateral intercostal artery perforator （LICAP） flap. Methods From January 2009 to June 2012, tissue defects in the axillary chest wall region of 8 patients aged from 13 to 22 years after microcystic LMs resections were closed using LICAP flap. Flap donor sites in all patients were closed primarily except one patient who underwent skin grafting. Before surgery, ultrasound and MRI examination were used to confirm the diagnosis and determine the scope and level of the abnormality for complete resection. Results All defects after microcystic LMs excision were successfully closed using LICAP flaps. The follow-up period ranged from 1 to 3 years （mean 2.1 years）. All flaps survived postoperatively. No recurrence occurred. Ultrasound and MRI also demonstrated flap survival without recurrence of microcystic LMs. No functional loss attributable to the LICAP flap harvest was identified. Conclusion Ultrasound and MRI are safe and accurate diagnostic imaging methods for the pre- and post-operative evaluation of the microcystic LMs in patients undergoing surgery. LICAP flap provides good coverage for the large defects and achieves acceptable morphology without functional deficits in donor sites. Related Articles | Metrics

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Carving-Assembling Method For Autologous Costal Cartilage Ear Framework Fabrication LIU Tun,GUO Fen,ZHOU Xu,HU Jintian,WANG Yue,QIAN Jin,XIE Yangchun,ZHANG Qingguo 2014, 10 (6):  332-334.  DOI: 10.3969/j.issn.1673-0364.2014.06.008 Abstract ( 418 )   Objective To explore a carving-assembling method to fabricate an autologous costal cartilage framework for auricular reconstruction. Methods From Jan. 2011 to Jan. 2012, 545 cases received auricular reconstruction. Among all the cases, 285 cases were treated with modified Nagata method, others with expanded retroauricular flap. Ear framework was fabricated in layers using splicing method with 6th to 8th costal cartilage. The framework includes helix, antihelix, scapha, triangular fossa, superior crus of antihelix, tragus, antitragus and concha. Results Six costal frameworks exposed due to local flaps necrosis. Because of infection, 1 case of framework was removed and re-implanted to the ipsilateral chest. All the patients were followed up for 6-12 months. Among them, 488 cases of reconstructed ears obtained vivid details with symmetric size and position, 80 cases of reconstructed ears had undesirable details. Nineteen cases lost to follow-up. Conclusion Fabricating tridimensional autologous costal cartilage framework by carving-assembling methods is safe, stable and practical. It could be an ideal method for auricular reconstruction. Related Articles | Metrics

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Surgical Reconstruction of Chronic Radiation-induced Ulcer in Chest Wall by Stages ZHOU Yaodong,GUAN Xin,ZHANG Yinxin,FEN Shaoqing,WANG Feng,LIANG Xi,LIANG Xiang 2014, 10 (6):  335-338.  DOI: 10.3969/j.issn.1673-0364.2014.06.009 Abstract ( 320 )   Objective To explore the surgical reconstruction of chronic radiation-induced ulcer in chest wall by stages. Methods From April 2010 to June 2013, 10 patients with huge chronic chest wall defect underwent chest wall reconstruction by stages. In stage Ⅰ, patients received debridement and greater omentum tamping; In stage Ⅱ, patients received surgical reconstruction by regional rotation flap, including pedicle latissimus dorsi myocutaneous flap and pectoralis major myocutaneous flap. The clinical efficacy and postoperative complications were observed. Results All the patients were followed up for 26-37 months （mean 25 months）. There were no operating death and local failure. One patient were suffered from subcutaneous infection which healed by debridement and drainage. No infection were observed in other patients. No paradoxical respiration occurred in all patients. Conclusion Stage therapeutics is effective in treating chronic radiation-induced ulcer and defect. Great omentum flap is useful for the repair of soft tissue defect, which is priority for defects caused by infection. Related Articles | Metrics

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Evaluation on Two Kinds of Cell-assisted Lipotransfer for Breast Augmentation WANG Lin,LU Yi,FU Mingang,HE Jiyin,HUANG yixiong,FAN Zhihong 2014, 10 (6):  339-342.  DOI: 10.3969/j.issn.1673-0364.2014.06.010 Abstract ( 303 )   Objective To evaluate the effect of two kinds of cell-assisted lipotransfer for breast augmentation. Methods Thirty patients accepted breast augmentation using cell-assisted lipotransfer （CAL）, and completed 12-month follow-up. According to the stromal vascular fraction （SVF） distilled from adipose tissue or aspirated liquid, the patients were divided into 2 groups：S group and L group. Flow-cytometry was used to detect the percentage of adipose-derived stem cells （ADSCs） in distilled SVF. The breast volumes and images were evaluated by MRI pre-operative, 6 and 12 months post-operative. The data of breast volume were marked and the resorption rate of transplanted adipose tissue was calculated. Results Averagely, the percentage of ADSCs in freshly distilled SVF in S group is 40.73% and 3.75% in L group. MRI showed that the breast volume 6-month post-operative and 12-month post-operative were both larger than that of preoperative in S group （P〈0.05）, while no significant difference between the breast volume 6-month post-operative and 12-month post-operative were observed （P〉0.05）, neither the resorption rate of 6 and 12-month post-operative （P〉0.05）, and the resorption rate of transplanted adipose tissue 12-month post-operative in S group was （51.83±15.28）%. However, there were no significant difference of breast volume among three time points in L group （P〉0.05）, neither the resorption rate of 6 and 12-month post-operative （P〉0.05）, and the resorption rate of transplanted adipose tissue 12-month post-operative in L group was （75.47±12.20）%. In addition, no calcification was seen in all MRI images but cysts were found in two cases of S group. Conclusion In process of breast augmentaion using cell-assissted lipotransfer, the distilled SVF from adipose tissue contains more ADSCs than that from aspirated liquid. The cell-assisted lipotransfer for breast augmentation is effective, safe, and the shrinkage stalls at 6-month post-operative. Related Articles | Metrics

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The Clinical Efficacy of Gastrocnemius Muscle Flap Combining with Vacuum Sealing Drainage in Treating Chronic Tibia Osteomyelitis with Soft Tissue Defect YANG Dai,FANG Bo,SONG Guixian,YAN Xinhai,SUN Shuhai,SHEN Yuzhen 2014, 10 (6):  343-345.  DOI: 10.3969/j.issn.1673-0364.2014.06.011 Abstract ( 259 )   Objective To explore the effect of gastrocnemius muscle flap combining vacuum sealing drainage （VSD） in the treatment of chronic osteomyelitis with soft tissue defect in tibia. Methods From Jan. 2010 to Dec. 2013, a total of 20 patients with chronic tibia osteomyelitis with soft tissue defect were analyzed retrospectively. The patients were divided into 2 groups, 8 cases were treated with traditional methods （control group） and other 12 cases were treated with gastrocnemius muscle flap combining VSD （experimental group）. Changing rate of CRP 72 hours post-operative, one-time cure rate, healing time and recurrence rate were assessed. Results All the indexes in experimental group were better than in control group （P＆lt;0.05）. Comparing with control group, the changing rate of CRP 72 hours post-operatively and one-time cure rate in experimental group were higher, while the healing time was lower. The recurrence rate of experimental group was 0%. Conclusion Gastrocnemius muscle flap combining with VSD is effective in treating chronic osteomyelitis with soft tissue defect in tibia with stable curative effect, quick recovery and low recurrence rate, and is worthy of promotion. Related Articles | Metrics

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The Short-term Prognosis and Safety Analysis of Modified Three-point Double Eyelid Plasty in Treating Blepharochalasis SHI Wenjuan,WU Suofa 2014, 10 (6):  346-354.  DOI: 10.3969/j.issn.1673-0364.2014.06.012 Abstract ( 203 )   Objective To explore the short-term effectiveness and safety of modified three-point double eyelid plasty in the treatment of blepharochalasis. Methods From March 2012 to March 2012, 74 patients with blepharochalasis were randomly divided into two groups by sampling. In control group （n=37）, the patients were treated with the traditional double eyelid plasty （embedding, incision or suture method）, while the patients were treated with the modified three-point double eyelid plasty in experimental group. The clinical curative effect were observed in the two groups. Results The post-operative recovery time in experimental group （35.23±6.56 days） was obviously shorter than in control group （79.81±13.24 days） （P〈0.05）. But there were no significant differences between two groups in primary healing rate and complication rate （ P〉0.05）;The degree of satisfaction in experimental group （94.59%） was significantly higher than in control group （78.38%） （ P〈0.05）. Conclusion The clinical curative effect of the modified three point double eyelid plasty is superior to the traditional methods in treating blepharochalasis with high safety index and short recovery time, is worthy of clinical application. Related Articles | Metrics

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Differentiation of Adipose-derived Stem Cells into Cardiomyocytes SHI Yuanyuan,YANG Xiangqun 2014, 10 (6):  351-354.  DOI: 10.3969/j.issn.1673-0364.2014.06.014 Abstract ( 392 )   The use of stem cells in the treatment of myocardial infarct has been strongly pursued in recent years. Due to their availability, feasibility, plasticity, and their potential to differentiate into cardiomyocytes at specific media, adipose-derived stem cells （ASCs） have become an ideal source of donor cells. A significant effort has gone into elucidating which factors and techniques influence this differentiation. The methods and mechanisms of differentiation of ASCs into cardiomyocytes were reviewed in this paper. Related Articles | Metrics

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Research Progress of Bone Morphogenetic Protein 2/7 Heterodimer GAO Tao,ZHOU Lulu,MIAO Chunlei,TANG Shengjian,LIU Fangjun 2014, 10 (6):  355-362.  DOI: 10.3969/j.issn.1673-0364.2014.06.015 Abstract ( 352 )   BMP2/7 heterodimer, a bone structure formation protein, has stronger osteogenesis inductive potential than its homodimer. So the study of BMP2/7 heterodimer has been one of the most important research directions in tissue engineering osteogenesis now. In this article, the osteo-inductive related experiments of BMP2/7 heterodimer were described, the differences of biological activity between heterodimer and homodimer were compared, and the mechanisms were overviewed. Related Articles | Metrics

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Overview and Research Progress of microRNA QI Bin,YU Limei,Zunyi Medical college, 2014, 10 (6):  357-359.  DOI: 10.3969/j.issn.1673-0364.2014.06.016 Abstract ( 702 )   The microRNA is only 20-22 nucleotides in length and can work as post-DNA transcription regulator for gene expression. It has an important regulating role in the life process such as cell aging, disease and so on. So microRNA has a huge potential for anti-aging and disease treatment. Nowadays it has become one of the most intense interests to the tissue engineering and other area of medicine. In this paper, the biogenesis, mechanism, relationship with cell aging and age-associated diseases and the clinical application of microRNA were reviewed. Related Articles | Metrics

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Pulsed Dye Laser Treatment in Infantile Hemangioma LV Dongze,MA Gang,LIN Xiaoxi 2014, 10 (6):  360-362.  DOI: 10.3969/j.issn.1673-0364.2014.06.017 Abstract ( 754 )   Infantile hemangioma （IH） is the most common benign tumor of infancy. Although it is self-limiting, its cosmetic problems and complications still has been the psychological concerns of the patients and parents. Pulsed dye laser （PDL） is the criterion standard for treating vascular lesions. It is controversial in treating IH, despite the 20 years＇ application. In this article, the current knowledge of the PDL treatment of IH was reviewed comprehensively and the new implications and directions for management of IH in the future were provided. Related Articles | Metrics

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Decompressing Peripheral Nerves of Lower Extremity in Patients with Diabetic Neuropathy for Preventing Foot Ulceration ZHANG Yang,ZHU Lian,LI Qingfeng 2014, 10 (6):  363-366.  DOI: 10.3969/j.issn.1673-0364.2014.06.018 Abstract ( 390 )   Diabetic foot ulceration can be closely associated with the lesion of peripheral nerves, which are more vulnerable to oppression in anatomical narrow places. Peripheral neurolysis has become a treatment for diabetic peripheral neuropathy through releasing the compressed nerves. It can not only improve the symptoms caused by diabetic peripheral neuropathy, but also reduce the incidence of foot ulceration. In this article, the basic scientific and clinical researches that preventing foot ulceration by decompressing peripheral nerves of lower extremity in patients with diabetic neuropathy were reviewed. Related Articles | Metrics