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Reconstruction of Larger Nasal Defects using Tip Divided-Scalping Forehead Flap 鼻尖分剥额瓣修复较大鼻部缺损
Pub Date : 2016-03-21 DOI: 10.4172/2157-7552.1000166
Y. Sowa, Toshiaki Numajiri
The scalping forehead flap, first reported by Converse in 1942 [1], is one of the best techniques for total and subtotal nasal reconstruction because this flap is extremely reliable and provides skin of good quality and colour-match [2]. We introduce the technique and refinements of nasal reconstruction for defects together with the nasal lining and the upper lip by using the divided distal part of the scalping forehead flap. This method provides adequate tissue for large and three-dimensional defects in a one-stage procedure.
1942年Converse首次报道的头皮前额皮瓣[1]是全鼻和次全鼻重建的最佳技术之一,因为这种皮瓣非常可靠,提供了良好的皮肤质量和颜色匹配[2]。本文介绍了利用头皮额瓣的远端分割部分修复鼻缺损及鼻衬和上唇的技术和改进。这种方法在一个阶段的程序中为大的三维缺陷提供了足够的组织。
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引用次数: 2
Osteochondral Regeneration of the Loading-bearing Site Using a Scaffold Free Three-dimensional Construct of Adipose Tissue-derived Mesenchymal Stem Cells in Pigs 利用无支架的脂肪组织源性间充质干细胞三维构建猪承重部位骨软骨再生
Pub Date : 2016-03-03 DOI: 10.4172/2157-7552.1000165
D. Murata, S. Tokunaga, Shizuka Akieda, K. Nakayama, Kentaroh Setoyama, M. Fujiki, K. Misumi
Background: Many surgical strategies for reconstruction of both bone and cartilage have ever been investigated to restore joint structure and function in the late stages of Osteoarthritis (OA). This study was designed to investigate the regeneration of articular cartilage and subchondral bone in the loading-bearing site using a three-dimensional (3D) construct of autologous adipose tissue-derived mesenchymal stem cells (AT-MSCs). Methods: A 3D construct consisting of approximately 1,920 spheroids each containing 5.0 × 104 AT-MSCs was implanted into an osteochondral defect (with a diameter of 6.8 mm and a depth of 6 mm) in the right femoral medial condyle in five adult mini-pigs. The contralateral (left femoral) defect was the control. At three and six months post-operatively, the defects were evaluated using both CT and MR imaging. The radiolucent volume (RV, mm3) of the defects was calculated based on the multiplanar reconstruction of the CT images. MR images and gross and histologic pathology features were scored using a modified-MOCART system and the ICRS system, respectively, at six months post-operatively. Results: The percentages of RVs at three and six months compared with those immediately after the surgeries were significantly decreased in the implanted defects compared with the control defects. The total scores of modified- MOCART system were also significantly increased in the implanted sites comparing to the controls. Although there were no statistical differences in the average of gross scores, the average histological scores were significantly higher in the implanted sites than in the control sites. Conclusion: This is the first report suggesting that implantation of a scaffold-free three dimensional construct of only AT-MSCs into the osteochondral defect regenerates the original cartilage and subchondral bone structures over six months post-operatively in the loading-bearing site of large animal.
背景:许多骨和软骨重建的手术策略已经被研究过,以恢复骨性关节炎(OA)晚期的关节结构和功能。本研究旨在利用自体脂肪组织源性间充质干细胞(AT-MSCs)的三维结构研究关节软骨和软骨下骨在负重部位的再生。方法:将约1,920个含5.0 × 104个AT-MSCs的球体三维构建体植入5只成年小型猪右股内侧髁骨软骨缺损(直径6.8 mm,深度6 mm)。对照组为对侧(左股)缺损。在术后3个月和6个月,使用CT和MR成像评估缺陷。基于CT图像的多平面重建计算缺陷的辐射透光体积(RV, mm3)。术后6个月,分别使用改良的mocart系统和ICRS系统对MR图像、大体和组织病理特征进行评分。结果:植入缺损与对照缺损相比,术后3个月和6个月的RVs百分比明显降低。改良MOCART系统在植入部位的总得分也明显高于对照组。虽然毛评分的平均值没有统计学差异,但植入部位的平均组织学评分明显高于对照部位。结论:这是首次报道将无支架的AT-MSCs三维构建体植入骨软骨缺损,可在术后6个月内在大型动物负重部位再生原有的软骨和软骨下骨结构。
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引用次数: 1
In Vitro Rooting and Acclimatization of Micropropagated Elite Sugarcane ( Saccharum officinarum L.)Genotypes - N52 and N53 优质甘蔗(Saccharum officinarum L.)的离体生根与驯化基因型- N52和N53
Pub Date : 2016-02-11 DOI: 10.4172/2157-7552.1000164
Melaku Tesfa, B. Admassu, K. Bantte
Availability of sufficient quantity and quality of sugarcane planting materials from conventional seed source is one of the major challenges in the Ethiopian sugar estates. To circumvent this challenge, tissue culture technology is found to be the best alternative for which in vitro propagation protocol is a key pre-request. Thus, the present study was aimed to optimize protocol for in vitro rooting and acclimatization of two elite sugarcane genotypes i.e., N52 and N53. Experiments were laid out in a completely randomized design with factorial treatment arrangements. Half strength MS liquid media supplemented with combination of Sucrose (0, 40, 50, 60 and 70 g/l) and NAA (0,3,5 and 7 mg/l) along with two sugarcane genotypes (N52, N53) were used for rooting while substrate containing sand, soil and farmyard manure in six different ratios (1:1:0, 1:1:1, 1:2:1, 2:1:1, 1:1:2 and 1:2:0) were used for acclimatization. With regard to in vitro rooting, ½ strength liquid MS medium + 50 g/l sucrose + 3 mg/l NAA induced the highest rooting (100%) with 23.5 ± 1.29 average root number per shoot and 4.95 cm ± 0.06 cm root length in genotype N52 while 5 mg/l NAA + 50 g/l sucrose induced the highest (100%) rooting response with an average of 21.76 ± 0.57 root number per shoot with 4.54 cm ± 0.06 cm root length in sugarcane genotype N53. In acclimatization, best survival rate (94% in N52 and 100% in N53) was achieved on substrate mixtures containing sand + soil in 1:1: ratios. Thus, it can be deduced that this protocol can be used successfully for in vitro rooting and acclimatization of these genotypes.
从传统种子来源获得足够数量和质量的甘蔗种植材料是埃塞俄比亚糖业面临的主要挑战之一。为了规避这一挑战,组织培养技术被认为是最好的选择,体外繁殖协议是一个关键的先决条件。因此,本研究旨在优化两个甘蔗优良基因型N52和N53的离体生根和驯化方案。实验采用完全随机设计,并采用因子处理安排。采用半强度MS液体培养基,分别添加蔗糖(0、40、50、60和70 g/l)和NAA(0、3、5和7 mg/l)以及两种甘蔗基因型(N52、N53)进行生根,并以6种不同比例(1:1:0、1:1:1、1:2:1、2:1:1、1:1:2和1:2:0)的沙、土和农家肥为基质进行驯化。关于体外加油,½强度液体介质女士+ 50 g / l蔗糖+ 3 mg / l NAA诱导生根最高(100%),23.5±1.29平均根数拍±0.06厘米和4.95厘米根长度在基因型N52 5 mg / l NAA + 50 g / l蔗糖诱导最高(100%)支持响应平均21.76±0.57根数量每拍摄±0.06厘米和4.54厘米根甘蔗基因型N53长度。驯化时,以1:1比例的砂土混合基质的成活率最高(N52为94%,N53为100%)。由此可以推断,该方案可以成功地用于这些基因型的离体生根和驯化。
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引用次数: 9
Mechanical Isolation of Human Endothelial Cells from Large Vessels for Flow Cytometry Immunophenotyping 人大血管内皮细胞机械分离的流式细胞术免疫分型研究
Pub Date : 2016-02-05 DOI: 10.4172/2157-7552.1000163
C. Torres, Rui Machado, M. Lima
Endothelial cells (EC) have important physiological functions, and they may also have a role in pathology. To better understand their role in health and disease, we must know very well their phenotype. Previous studies have identified and characterized EC mainly by immunohistochemistry, but there are also some studies using flow cytometry (FCM) after exposing these cells to enzymatic digestion, either to isolate and/ or to detach them from the vessel wall. However, it is well known that enzymatic treatment can cause deleterious effects on cell surface receptors, then influencing the antibody-antigen reaction. We describe a simple and cheap mechanical method to isolate EC from human vessels, avoiding alterations in the expression of cell surface receptors caused by the use of enzymes, and we tested it using FCM. With this method we were able to obtain fresh EC that were identified by FCM as a well-defined cluster of CD45-CD146+brightCD31+bright cells. This approach can be used in the future to isolate EC for further immunophenotypic characterization and for ex-vivo functional studies, as well as to test the effect of different stimuli, including pharmacological drugs.
内皮细胞(EC)具有重要的生理功能,也可能在病理中发挥作用。为了更好地理解它们在健康和疾病中的作用,我们必须很好地了解它们的表型。先前的研究主要通过免疫组织化学鉴定和表征EC,但也有一些研究在将这些细胞暴露于酶消化后使用流式细胞术(FCM)分离和/或从血管壁分离它们。然而,众所周知,酶处理会对细胞表面受体产生有害影响,从而影响抗体-抗原反应。我们描述了一种简单廉价的从人血管中分离EC的机械方法,避免了酶引起的细胞表面受体表达的改变,并使用流式细胞仪进行了测试。通过这种方法,我们能够获得新鲜的EC,通过流式细胞仪鉴定为一个定义明确的CD45-CD146+brightCD31+bright细胞簇。这种方法可以在未来用于分离EC,以进一步进行免疫表型表征和离体功能研究,以及测试不同刺激的效果,包括药理学药物。
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引用次数: 2
Atelocollagen Enhanced Osteogenesis in a Geometric Structured Beta-TCP Scaffold by VEGF Induction 血管内皮生长因子诱导胶原蛋白促进几何结构β - tcp支架成骨
Pub Date : 2016-01-18 DOI: 10.4172/2157-7552.1000162
K. Yagami, Sunao Sadaoka, Hiroshi Nakamura, S. Komatsu, J. Onodera, Masahiko Suzuki, Y. Kuboki
Abstract In order to establish the convertibility of a host for bone augmentation, we herein developed a new honeycombshaped β-tricalcium phosphate (37H) using atelocollagen as a scaffold, which exhibited unique geometric properties for in vitro and in vivo studies. Human mesenchymal stem cells (MSC) were cultured with 37H or atelocollagen-coated honeycomb-shaped β-tricalcium phosphate (Col37H), and their osteoblastic differentiation was then analyzed. Atelocollagen promoted cell adhesion and formation of vessel-like structures in the tunnels of scaffolds of cultured MSC. The mRNA expression levels of type I collagen, osteocalcin, and VEGF were greater in MSC cultured with Col37H than with 37H. Bone generation with Col37H in the rat calvaria was greater than with 37H, and this was attributed to early vessel construction. A large number of blood vessels invaded tunnels from the periosteum and existing bone surface. A strong VEGF signal was detected immediately before the new bone surface in the tunnels of Col37H. These results indicate that the addition of atelocollagen to Col37H has potential in the construction of functional artificial bone.
为了建立骨增强宿主的可转换性,我们在此开发了一种新型蜂窝状的β-磷酸三钙(37H),该支架以胶原为支架,在体外和体内研究中均表现出独特的几何特性。用37H或胶原包被蜂窝状β-磷酸三钙(Col37H)培养人间充质干细胞(MSC),观察其成骨分化情况。胶原蛋白促进细胞黏附和血管样结构的形成。Col37H培养的MSC中I型胶原、骨钙素和VEGF mRNA表达水平高于37H培养的MSC。大鼠颅骨中Col37H的骨生成大于37H,这归因于早期血管构建。大量血管从骨膜和现有骨表面侵入隧道。在Col37H的隧道中,在新骨表面形成之前检测到强烈的VEGF信号。这些结果表明,在Col37H中添加间胶原具有构建功能性人工骨的潜力。
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引用次数: 1
Synergy between rhBMP-2 and IKK-Inhibitor PS-1145 Delivered via a Porous Biodegradable Polymer Implant rhBMP-2和ikk抑制剂PS-1145的协同作用通过多孔可生物降解聚合物植入物传递
Pub Date : 2016-01-01 DOI: 10.4172/2157-7552.S1-003
David Carr, N. Y. Yu, J. Fitzpatrick, L. Peacock, K. Mikulec, A. Ruys, J. Cooper-White, D. Little, A. Schindeler
Critical-sized bone defects, whether caused by congenital malformation, tumor resection, trauma, or implant loosening, remain a major challenge for orthopaedic management. In this study we describe a bone tissue engineering approach in mice for the co-delivery of recombinant human Bone Morphogenetic Protein-2 (rhBMP-2) and the IKK inhibitor PS-1145. Scaffold implants were manufactured from poly(lactide-co-glycolide)(PLGA) by Thermally-Induced Phase Separation (TIPS), with rhBMP-2 (10 μg) and the IKK inhibitor PS-1145 (0 μg, 40 μg or 80 μg) incorporated into the polymer. These scaffolds were then surgically implanted into the hind limb muscle of C57BL6/J mice. One group of mice also received systemic 50 mg/kg PS-1145 (days 11-20). Specimens were harvested at week 3 for X-ray and microCT analyses and descriptive histology. Local and systemic delivery PS-1145 both significantly increased the net rhBMP-2 induced bone at 3 weeks. A maximal response was seen with the 40 μg PS-1145 group, although there was no significant difference between the 40 μg and 80 μg PS-1145 regimens. No local cytotoxicity was seen with either dose of PS-1145. In summary, local co-delivery of rhBMP-2 and PS-1145 via a porous PLGA scaffold represents a new tissue engineering approach for maintaining new bone in an unloaded environment.
无论是由先天性畸形、肿瘤切除、创伤还是植入物松动引起的严重骨缺损,仍然是骨科治疗的主要挑战。在这项研究中,我们描述了一种骨组织工程方法,用于在小鼠中共同递送重组人骨形态发生蛋白2 (rhBMP-2)和IKK抑制剂PS-1145。采用热诱导相分离(TIPS)法制备PLGA支架植入物,将rhBMP-2 (10 μg)和IKK抑制剂PS-1145 (0 μg, 40 μg或80 μg)掺入聚合物中。然后将这些支架手术植入C57BL6/J小鼠后肢肌肉。一组小鼠同时全身注射50 mg/kg PS-1145(第11-20天)。标本于第3周采集,用于x射线和显微ct分析和描述性组织学。局部和全身给药PS-1145均在3周时显著增加rhBMP-2诱导的净骨。40 μg PS-1145组反应最大,但40 μg PS-1145组与80 μg PS-1145组无显著性差异。两种剂量的PS-1145均未见局部细胞毒性。总之,通过多孔PLGA支架局部共同递送rhBMP-2和PS-1145代表了一种新的组织工程方法,用于在卸载环境中维持新骨。
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引用次数: 3
Clinical Outcomes of Esthetic and Functional Rehabilitation with Dental Implants in Patients with Alveolar Cleft 牙槽裂患者种植体的美观和功能康复的临床效果
Pub Date : 2015-12-28 DOI: 10.4172/2157-7552.1000160
Hidemi Nakata, S. Kuroda, N. Tachikawa, Munemitsu Miyasaka, K. Yoneda, H. Kondo, S. Kasugai
Background and aim: Alveolar cleft is notable congenital deformity in the oral and maxillofacial region. And missing incisor is often associated with the cleft site. Owing to recent advances in bone grafting techniques, dental implant treatment has become an appreciable method of restoring the edentulous space at the cleft site in addition to conventional methods using bridges or dentures. In this study, we investigated the risk factors related to esthetic outcomes of implant treatment at alveolar cleft sites. Materials and methods: A total of 13 patients treated with dental implants for missing teeth associated with an alveolar cleft were examined. The patients’ gender, cleft type, and number of dental implants were recorded. The ages of patients at bone grafting for cleft closure, additional bone grafting prior to dental implant surgery, and dental implant placement were also investigated. Seven risk factors were assessed to evaluate the relation to esthetic outcomes. Results: Five to thirteen years after the final prostheses were delivered, neither marginal bone resorption nor loss of dental implants were found. The esthetic outcomes differed among patients according to several risk factors. Discussion and conclusion: Oral rehabilitation with dental implants represented a promising treatment for alveolar cleft sites when bone graft was appropriate. However, the esthetic outcomes of this treatment depend upon multiple risk factors. Type of Clefts r-s l-s Bilateral Total Male 1 3 1 5 Female 0 6 2 8 Total 1 9 3 13 r-s: right side l-s: left side Table 1: Description of the 13 patients evaluated in the present study. Journal of Tissue Science & Engineering J o u r n a l o f T iss ue S cience &ngine e r i n g
背景与目的:牙槽裂是口腔颌面部显著的先天性畸形。切牙缺失通常与唇裂部位有关。由于近年来骨移植技术的进步,种植牙治疗已经成为除了使用桥或假牙的传统方法之外,修复裂隙部位无牙空间的一种很好的方法。在这项研究中,我们调查了与牙槽裂部位种植体治疗美学结果相关的危险因素。材料与方法:对13例因牙槽裂缺失而行种植牙治疗的患者进行检查。记录患者性别、裂型、种植牙数量。我们还调查了腭裂闭合植骨患者的年龄、种植牙手术前的额外植骨以及种植牙的放置情况。对7个危险因素进行评估,以评估其与审美结果的关系。结果:术后5 ~ 13年未见种植体边缘骨吸收和丢失。根据几个危险因素,患者的审美结果有所不同。讨论与结论:当骨移植合适时,种植牙的口腔康复是一种很有前途的治疗牙槽骨裂的方法。然而,这种治疗的美观效果取决于多种危险因素。唇裂类型r-s l-s双侧总数男性13 1 5女性0 6 2 8总数1 9 3 13 r-s:右侧l-s:左侧表1:本研究评估的13例患者的描述。组织科学与工程学报[J] .中国生物医学工程学报
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引用次数: 0
Methodological Consideration of Various Intraosseous and Heterotopic Bone Grafts Implantation in Animal Models 不同骨内和异位骨移植动物模型的方法学思考
Pub Date : 2015-12-17 DOI: 10.4172/2157-7552.1000161
Weiguo Xu, G. Spilker, C. Wein
Bone fracture healing is a complex process including inflammation, repair and remodeling. Bone grafts or substitute are widely accepted to treat impaired healing. Newly developed bone substitutes must undergo in vitro and in vivo testing before clinical application. All kinds of intraosseous and heterotopic implant models in small and/ or large animals are used for different bone substitutes. Orthotopic implant at models is used for evaluation of graft materials. Based on the statistic of literatures, we find small animals should be used first before large animal as osseous defect models. Rabbit and rat are the commonly chosen animals, while femur and calvaria are the most implanted anatomic sites. Critical size defect models are useful as bone defect model, but vary considerably between animals. Typical heterotopic ossification after implantation of bone substitute is found almost in all species of animals. No bone formation is found after subcutaneous implantation of bone granule grafts in small animal, but in large animal. In contrast, bone block grafts show a distinguished result of bone formation in small animals as well as in large animals. This article reviews currently animal bone defect models and anatomic implant site for bone graft, gives a recommendation for the future research.
骨折愈合是一个复杂的过程,包括炎症、修复和重塑。骨移植或骨替代物被广泛接受用于治疗愈合受损。新开发的骨替代物在临床应用前必须经过体外和体内试验。各种小型和/或大型动物的骨内和异位种植体模型用于不同的骨代用品。原位种植体模型用于评估移植材料。通过文献统计,我们发现小动物作为骨缺损模型应优先于大动物。兔和大鼠是常用的植入动物,股骨和颅骨是植入最多的解剖部位。临界尺寸缺陷模型是有用的骨缺陷模型,但在动物之间差异很大。骨替代物植入后的典型异位骨化几乎在所有物种的动物中都存在。小动物皮下植入骨颗粒后未见骨形成,而大型动物皮下植入骨颗粒后未见骨形成。相比之下,骨块移植在小动物和大动物中都显示出不同的骨形成结果。本文综述了目前动物骨缺损模型和骨移植的解剖植入部位,并对今后的研究提出了建议。
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引用次数: 1
Fabrication, Mercury Intrusion Porosimetry Characterization and In Vitro Qualitative Analysis of Biocompatibility of Various Porosities Polycaprolactone Scaffolds 不同孔隙度聚己内酯支架的制备、压汞孔法表征及体外生物相容性定性分析
Pub Date : 2015-12-02 DOI: 10.4172/2157-7552.1000159
T. Limongi, A. Giugni, H. Tan, Ebtihaj M Bukhari, B. Torre, M. Allione, M. Marini, L. Tirinato, G. Das, M. Moretti, A. Falqui, E. Fabrizio
In order to develop surfaces with improved cell culture biocompatibility, we optimized a solvent-casting and particulate-leaching fabrication technique to create porous three-dimensional polycaprolactone scaffolds. These biocompatible porous surfaces were realized by means of NaCl particles as porogen; salt leaching by immersion in distilled water created porosity and pore interconnectivity in the material. Scanning electron microscopy and mercury intrusion porosimetry were used for the measurement of porosity, pore size distribution, permeability and compressibility. To evaluate scaffold biocompatibility, fibroblasts were cultured on the porous surfaces and confocal immunofluorescence characterization indicated that they were effective for in vitro cell culture and practical tissue engineering applications.
为了开发具有更好的细胞培养生物相容性的表面,我们优化了溶剂铸造和颗粒浸出制造技术,以制造多孔的三维聚己内酯支架。这些生物相容性多孔表面是通过NaCl颗粒作为孔隙剂实现的;浸没在蒸馏水中的盐浸出在材料中产生孔隙和孔隙互连性。采用扫描电镜和压汞法测定孔隙度、孔径分布、渗透率和压缩性。为了评估支架的生物相容性,在多孔表面培养成纤维细胞,共聚焦免疫荧光表征表明它们在体外细胞培养和实际组织工程应用中是有效的。
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引用次数: 3
The Application of Stem Cell Based Tissue Engineering in Spinal Cord Injury Repair. 干细胞组织工程在脊髓损伤修复中的应用。
Pub Date : 2015-11-24 DOI: 10.4172/2157-7552.1000158
Wanting Niu, Xiang Zeng
Spinal Cord Injury (SCI) results in the permanent functional impairment, leading to monoplegia, paraplegia or tetraplegia with tremendous social and economic burden. The intrinsic repair mechanism has been proven to be insufficient. The complex pathophysiology after injury imposes enormous challenging to functional recovery, given the most advanced medical intervention nowadays. Therefore, the development of effective therapeutic strategy for spinal cord injury management is in great need. Here we review a stem cell based tissue engineering approach under preclinical or clinical development for spinal cord injury, with a focus on promoting functional recovery after SCI, aiming to provide some beneficial suggestion on stem cell based tissue engineering design.
脊髓损伤(Spinal Cord Injury, SCI)可导致永久性的功能损害,导致单侧瘫痪、截瘫或四肢瘫痪,给患者带来巨大的社会和经济负担。内在修复机制已被证明是不足的。在当今最先进的医疗干预手段下,损伤后复杂的病理生理对功能恢复提出了巨大的挑战。因此,迫切需要制定有效的脊髓损伤治疗策略。本文综述了基于干细胞的组织工程治疗脊髓损伤的临床前和临床研究进展,重点介绍了促进脊髓损伤后功能恢复的方法,旨在为基于干细胞的组织工程设计提供一些有益的建议。
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引用次数: 2
期刊
Journal of Tissue Science and Engineering
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