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1.
Design and characterization of a tissue‐engineered bilayer scaffold for osteochondral tissue repair
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Paolo Giannoni Erica Lazzarini Luca Ceseracciu Alberto C. Barone Rodolfo Quarto Silvia Scaglione 《Journal of tissue engineering and regenerative medicine》2015,9(10):1182-1192
Treatment of full‐thickness cartilage defects relies on osteochondral bilayer grafts, which mimic the microenvironment and structure of the two affected tissues: articular cartilage and subchondral bone. However, the integrity and stability of the grafts are hampered by the presence of a weak interphase, generated by the layering processes of scaffold manufacturing. We describe here the design and development of a bilayer monolithic osteochondral graft, avoiding delamination of the two distinct layers but preserving the cues for selective generation of cartilage and bone. A highly porous polycaprolactone‐based graft was obtained by combining solvent casting/particulate leaching techniques. Pore structure and interconnections were designed to favour in vivo vascularization only at the bony layer. Hydroxyapatite granules were added as bioactive signals at the site of bone regeneration. Unconfined compressive tests displayed optimal elastic properties and low residual deformation of the graft after unloading (< 3%). The structural integrity of the graft was successfully validated by tension fracture tests, revealing high resistance to delamination, since fractures were never displayed at the interface of the layers (n = 8). Ectopic implantation of grafts in nude mice, after seeding with bovine trabecular bone‐derived mesenchymal stem cells and bovine articular chondrocytes, resulted in thick areas of mature bone surrounding ceramic granules within the bony layer, and a cartilaginous alcianophilic matrix in the chondral layer. Vascularization was mostly observed in the bony layer, with a statistically significant higher blood vessel density and mean area. Thus, the easily generated osteochondral scaffolds, since they are mechanically and biologically functional, are suitable for tissue‐engineering applications for cartilage repair. Copyright © 2012 John Wiley & Sons, Ltd. 相似文献
2.
Tianzheng Deng Jing Lv Jianliang Pang Bing Liu Jie Ke 《Journal of tissue engineering and regenerative medicine》2014,8(7):546-556
In this study, a novel three‐dimensional (3D) heterogeneous/bilayered scaffold was constructed to repair large defects in rabbit joints. The scaffold includes two distinct but integrated layers corresponding to the cartilage and bone components. The upper layer consists of gelatin, chondroitin sulphate and sodium hyaluronate (GCH), and the lower layer consists of gelatin and ceramic bovine bone (GCBB). The two form a 3D bilayered scaffold (GCH–GCBB), which mimics the natural osteochondral matrix for use as a scaffold for osteochondral tissue engineering. The purpose of this study was to evaluate the efficacy of this novel scaffold, combined with chondrocytes and bone marrow stem cells (BMSCs) to repair large defects in rabbit joints. Thirty‐six large defects in rabbit femoral condyles were created; 12 defects were treated with the same scaffold combined with cells (group A); another 12 defects were treated with cell‐free scaffolds (group B); the others were untreated (group C). At 6 and 12 weeks, in group A hyaline‐like cartilage formation could be observed by histological examination; the newly formed cartilage, which stained for type II collagen, was detected by RT–PCR at high‐level expression. Most of the GCBB was replaced by bone, while little remained in the underlying cartilage. At 36 weeks, GCBB was completely resorbed and a tidemark was observed in some areas. In contrast, groups B and C showed no cartilage formation but a great amount of fibrous tissue, with only a little bone formation. In summary, this study demonstrated that a novel scaffold, comprising a top layer of GCH, having mechanical properties comparable to native cartilage, and a bottom layer composed of GCBB, could be used to repair large osteochondral defects in joints. Copyright © 2012 John Wiley & Sons, Ltd. 相似文献
3.
In vitro and in vivo co‐culture of chondrocytes and bone marrow stem cells in photocrosslinked PCL–PEG–PCL hydrogels enhances cartilage formation
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Shu‐Rui Yang Tsai‐Yu Lin Sydney Peng Yu‐Shiang Peng Ming‐Huei Cheng I‐Ming Chu 《Journal of tissue engineering and regenerative medicine》2016,10(10):E485-E496
Chondrocytes (CH) and bone marrow stem cells (BMSCs) are sources that can be used in cartilage tissue engineering. Co‐culture of CHs and BMSCs is a promising strategy for promoting chondrogenic differentiation. In this study, articular CHs and BMSCs were encapsulated in PCL–PEG–PCL photocrosslinked hydrogels for 4 weeks. Various ratios of CH:BMSC co‐cultures were investigated to identify the optimal ratio for cartilage formation. The results thus obtained revealed that co‐culturing CHs and BMSCs in hydrogels provides an appropriate in vitro microenvironment for chondrogenic differentiation and cartilage matrix production. Co‐culture with a 1:4 CH:BMSC ratio significantly increased the synthesis of GAGs and collagen. In vivo cartilage regeneration was evaluated using a co‐culture system in rabbit models. The co‐culture system exhibited a hyaline chondrocyte phenotype with excellent regeneration, resembling the morphology of native cartilage. This finding suggests that the co‐culture of these two cell types promotes cartilage regeneration and that the system, including the hydrogel scaffold, has potential in cartilage tissue engineering. Copyright © 2013 John Wiley & Sons, Ltd. 相似文献
4.
Manuela E. Gomes Carlos A. Viegas Jorge T. Azevedo Isabel R. Dias Fernando M. Guzón Rui L. Reis 《Journal of tissue engineering and regenerative medicine》2011,5(1):41-49
This study aims to assess the in vivo performance of cell–scaffold constructs composed of goat marrow stromal cells (GBMCs) and SPCL (a blend of starch with polycaprolactone) fibre mesh scaffolds at different stages of development, using an autologous model. GBMCs from iliac crests were seeded onto SPCL scaffolds and in vitro cultured for 1 and 7 days in osteogenic medium. After 1 and 7 days, the constructs were characterized for proliferation and initial osteoblastic expression by alkaline phosphatase (ALP) activity. Scanning electron microscopy analysis was performed to investigate cellular morphology and adhesion to SPCL scaffolds. Non‐critical defects (diameter 6 mm, depth 3 mm) were drilled in the posterior femurs of four adult goats from which bone marrow and serum had been collected previously. Drill defects alone and defects filled with scaffolds without cells were used as controls. After implantation, intravital fluorescence markers, xylenol orange, calcein green and tetracycline, were injected subcutaneously after 2, 4 and 6 weeks, respectively, for bone formation and mineralization monitoring. Subsequently, samples were stained with Lévai–Laczkó for bone formation and histomorphometric analysis. GBMCs adhered and proliferated on SPCL scaffolds and an initial differentiation into pre‐osteoblasts was detected by an increasing level of ALP activity with the culture time. In vivo experiments indicated that bone neoformation occurred in all femoral defects. The results obtained provided important information about the performance of SPCL–GBMC constructs in an orthotopic goat model that enabled future studies to be designed to investigate in vivo the functionality of SPCL–GBMC constructs in more complex models, viz. critical sized defects, and to evaluate the influence of in vitro cultured autologous cells in the healing and bone regenerative process. Copyright © 2010 John Wiley & Sons, Ltd. 相似文献
5.
Biomimetic scaffolds and dynamic compression enhance the properties of chondrocyte‐ and MSC‐based tissue‐engineered cartilage
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Nopporn Sawatjui Temduang Limpaiboon Karsten Schrobback Travis Klein 《Journal of tissue engineering and regenerative medicine》2018,12(5):1220-1229
Adult chondrocytes are surrounded by a protein‐ and glycosaminoglycan‐rich extracellular matrix and are subjected to dynamic mechanical compression during daily activities. The extracellular matrix and mechanical stimuli play an important role in chondrocyte biosynthesis and homeostasis. In this study, we aimed to develop scaffold and compressive loading conditions that mimic the native cartilage micro‐environment and enable enhanced chondrogenesis for tissue engineering applications. Towards this aim, we fabricated porous scaffolds based on silk fibroin (SF) and SF with gelatin/chondroitin sulfate/hyaluronate (SF‐GCH), seeded the scaffolds with either human bone marrow mesenchymal stromal cells (BM‐MSCs) or chondrocytes, and evaluated their performance with and without dynamic compression. Human chondrocytes derived from osteoarthritic joints and BM‐MSCs were seeded in scaffolds, precultured for 1 week, and subjected to compression with 10% dynamic strain at 1 Hz, 1 hr/day for 2 weeks. When dynamic compression was applied, chondrocytes significantly increased expression of aggrecan (ACAN) and collagen X (COL10A1) up to fivefold higher than free‐swelling controls. In addition, dynamic compression dramatically improved the chondrogenesis and chondrocyte biosynthesis cultured in both SF and SF‐GCH scaffolds evidenced by glycosaminoglycan (GAG) content, GAG/DNA ratio, and immunostaining of collagen type II and aggrecan. However, both chondrocytes and BM‐MSCs cultured in SF‐GCH scaffolds under dynamic compression showed higher GAG content and compressive modulus than those in SF scaffolds. In conclusion, the micro‐environment provided by SF‐GCH scaffolds and dynamic compression enhances chondrocyte biosynthesis and matrix accumulation, indicating their potential for cartilage tissue engineering applications. 相似文献
6.
F. W. Janssen R. van Dijkhuizen‐Radersma A. Van Oorschot J. Oostra J. D. de Bruijn C. A. Van Blitterswijk 《Journal of tissue engineering and regenerative medicine》2010,4(1):12-24
The aim of this study was to evaluate a semi‐automated perfusion bioreactor system for the production of clinically relevant amounts of human tissue‐engineered bone. Human bone marrow stromal cells (hBMSCs) of eight donors were dynamically seeded and proliferated in a perfusion bioreactor system in clinically relevant volumes (10 cm3) of macroporous biphasic calcium phosphate scaffolds (BCP particles, 2–6 mm). Cell load and distribution were shown using methylene blue staining. MTT staining was used to demonstrate viability of the present cells. After 20 days of cultivation, the particles were covered with a homogeneous layer of viable cells. Online oxygen measurements confirmed the proliferation of hBMSCs in the bioreactor. After 20 days of cultivation, the hybrid constructs became interconnected and a dense layer of extracellular matrix was present, as visualized by scanning electron microscopy (SEM). Furthermore, the hBMSCs showed differentiation towards the osteogenic lineage as was indicated by collagen type I production and alkaline phosphatase (ALP) expression. We observed no significant differences in osteogenic gene expression profiles between static and dynamic conditions like ALP, BMP2, Id1, Id2, Smad6, collagen type I, osteocalcin, osteonectin and S100A4. For the donors that showed bone formation, dynamically cultured hybrid constructs showed the same amount of bone as the statically cultured hybrid constructs. Based on these results, we conclude that a semi‐automated perfusion bioreactor system is capable of producing clinically relevant and viable amounts of human tissue‐engineered bone that exhibit bone‐forming potential after implantation in nude mice. Copyright © 2009 John Wiley & Sons, Ltd. 相似文献
7.
Suzanne Tabbaa Karen J. L. Burg 《Journal of tissue engineering and regenerative medicine》2015,9(4):469-472
The major limitation of large tissue‐engineered constructs used for bone regeneration is the lack of vasculature and, therefore, lack of transport of essential nutrients, chemical factors and progenitor cells. Research approaches to improve the transport properties of large scaffolds focus on using angiogenic factors and vasculogenic cells to create new vasculature; however, the slow rate of vessel formation and reliance on vessel self‐assembly in these approaches is problematic. In this study, a novel approach has been proposed, using proprietary engineered ‘wicking’ fibres of non‐circular cross‐section that provide highly efficient transport for fluid and cells. The effect of wicking fibres on the movement of fluorescein isothiocyanate (FITC)‐conjugated protein in a three‐dimensional (3D) hydrogel system was analysed. The results indicated that the rate of diffusion of the fluorescent protein was greatly enhanced in hydrogels that contained wicking fibres in comparison to those that did not. The movement of progenitor cells along wicking fibres and round fibres was assessed. This study demonstrated that wicking fibres enhance the movement of critical growth factors and progenitor cells central for bone regeneration. The results suggested that the incorporation of wicking fibres into large tissue‐engineered constructs may improve the transport of growth factors and progenitor cells essential for bone formation. Copyright © 2014 John Wiley & Sons, Ltd. 相似文献
8.
Cultivation of agarose‐based microfluidic hydrogel promotes the development of large,full‐thickness,tissue‐engineered articular cartilage constructs
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Stephen M. Goldman Gilda A. Barabino 《Journal of tissue engineering and regenerative medicine》2017,11(2):572-581
The fabrication of tissue‐engineered constructs of clinically relevant sizes continues to be plagued by poor nutrient transport to the interior of the construct. Consequences of poor mass transfer to the construct core include large gradients in cell viability and matrix deposition, as well as inadequate mechanical functionality. Prior literature has shown that embedded microfluidic channels offer the potential to control the spatial and temporal presentation of hydrodynamic and chemical cues within the developing tissue construct toward improved mass transfer. The current state of the art in microfluidic constructs, however, has fallen short of achieving sufficient thickness and robustness of constructs for further development towards translation. Towards this goal, we designed a microfluidic tissue construct and established bioprocessing conditions to meet nutrient transport requirements of a large, full‐thickness, articular cartilage construct over a 2 week culture period. Our microfluidic constructs of 2.5 and 5 mm thicknesses showed enhanced cell proliferation relative to statically cultured constructs. These constructs, which are both thick and robust to culture periods of sufficient length to support extracellular matrix development, represent an important improvement over previously reported constructs which were thinner and lacking in extracellular matrix (most likely attributable to too‐short culture periods). Copyright © 2014 John Wiley & Sons, Ltd. 相似文献
9.
Biocompatibility evaluation of tissue‐engineered valved venous conduit by reseeding autologous bone marrow‐derived endothelial progenitor cells and multipotent adult progenitor cells into heterogeneous decellularized venous matrix
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Jian‐ming Yuan Zhen Liu Yong‐zhen Zhang Rui‐shan Dang Xiang‐qun Yang Xi Zhang Chuan‐sen Zhang 《Journal of tissue engineering and regenerative medicine》2016,10(12):982-988
Clinical treatment of chronic deep venous insufficiency remains difficult despite the availability of various therapies. Previous experimental efforts have demonstrated that the tissue‐engineered valvedvenous conduit (TEVV) is a promising option to replace the damaged venous valve. The aim of the present study was to evaluate the TEVV by reseeding bone marrow‐derived endothelial progenitor cells and multipotent adult progenitor cells into acellular matrix according to International Standard ISO10993, and to clarify their interactions with blood, the local effect after implantation both in vitro and vivo, and immunogenicity. The results showed that the 2‐cm long TEVV did not cause haemolysis in vitro and remained patent without thrombosis formation in vivo. However, the luminal surface of TEVV was partially covered by multilayer cells. Compared with the native ovine femoral vein segment, the TEVV beneath the mouse skin produced significant mononuclear cell infiltration, with serum interleukin‐6 and tumour necrosis factor‐α similar to normal. The TEVV maintained its structural integrity, while the native ovine femoral vein segments fell apart at postoperative week nine. The TEVV implantation did not change serum immunoglobulin G. In addition, the seeds and extracts of the scaffold did not affect the proliferation of mouse lymphocytes. These findings suggest that the histocompatibility, haemocompatibility and immunogenicity of this TEVV are acceptable owing to complete removal of the cellular components of autologous seeds and residues of chemical regents, thus providing an experimental basis for further clinical translation. Copyright © 2014 John Wiley & Sons, Ltd. 相似文献
10.
Three‐dimensional assembly of tissue‐engineered cartilage constructs results in cartilaginous tissue formation without retainment of zonal characteristics
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W. Schuurman E. B. Harimulyo D. Gawlitta T. B. F. Woodfield W. J. A. Dhert P. R. van Weeren J. Malda 《Journal of tissue engineering and regenerative medicine》2016,10(4):315-324
Articular cartilage has limited regenerative capabilities. Chondrocytes from different layers of cartilage have specific properties, and regenerative approaches using zonal chondrocytes may yield better replication of the architecture of native cartilage than when using a single cell population. To obtain high seeding efficiency while still mimicking zonal architecture, cell pellets of expanded deep zone and superficial zone equine chondrocytes were seeded and cultured in two layers on poly(ethylene glycol)‐terephthalate–poly(butylene terephthalate) (PEGT–PBT) scaffolds. Scaffolds seeded with cell pellets consisting of a 1:1 mixture of both cell sources served as controls. Parallel to this, pellets of superficial or deep zone chondrocytes, and combinations of the two cell populations, were cultured without the scaffold. Pellet cultures of zonal chondrocytes in scaffolds resulted in a high seeding efficiency and abundant cartilaginous tissue formation, containing collagen type II and glycosaminoglycans (GAGs) in all groups, irrespective of the donor (n = 3), zonal population or stratified scaffold‐seeding approach used. However, whereas total GAG production was similar, the constructs retained significantly more GAG compared to pellet cultures, in which a high percentage of the produced GAGs were secreted into the culture medium. Immunohistochemistry for zonal markers did not show any differences between the conditions. We conclude that spatially defined pellet culture in 3D scaffolds is associated with high seeding efficiency and supports cartilaginous tissue formation, but did not result in the maintenance or restoration of the original zonal phenotype. The use of pellet‐assembled constructs leads to a better retainment of newly produced GAGs than the use of pellet cultures alone. Copyright © 2013 John Wiley & Sons, Ltd. 相似文献
11.
Rohin K. Iyer Jane Chui Milica Radisic 《Journal of tissue engineering and regenerative medicine》2009,3(3):196-207
Cardiac tissue engineering aims to create myocardial patches for repair of defective or damaged native heart muscle. The inclusion of non‐myocytes in engineered cardiac tissues has been shown to improve the properties of cardiac tissue compared to tissues engineered from enriched populations of myocytes alone. While attempts have been made to mix non‐myocytes (fibroblasts, endothelial cells) with cardiomyocytes, very little is understood about how the tissue properties are affected by varying the respective ratios of the three cell types and how these cells assemble into functional tissues with time. The goal of this study was to investigate the effects of modulating the ratios of the three cell types and to spatially and temporally track cardiac tri‐cultures of cells. Primary neonatal cardiac fibroblasts and D4T endothelial cells were incubated in 5 µM CellTracker? green dye and CellTracker? red dye, respectively, while neonatal cardiomyocytes were labelled with 20 µg/mL DAPI. The non‐myocytes were seeded either sequentially (pre‐culture) or simultaneously (tri‐culture) in Matrigel‐coated microchannels and allowed to form organoids, as in our previous studies. We also varied the seeding percentage of cardiomyocytes while keeping the total cell number constant in an attempt to improve the functional properties of the organoids. Organoids were imaged on days 1 and 4. Endothelial cells were seen to aggregate into clusters when simultaneously tri‐cultured with myocytes and fibroblasts, while pre‐cultures contained elongated cells. Functional properties of organoids were improved by increasing the seeding percentage of enriched cardiomyocytes from 40% to 80%. Copyright © 2009 John Wiley & Sons, Ltd. 相似文献
12.
Bin Hu Jun‐Lin Ren Jun‐Rui Zhang Qin Ma Yan‐Pu Liu Tian‐Qiu Mao 《Journal of tissue engineering and regenerative medicine》2010,4(2):105-114
Direct intra‐articular injection of mesenchymal stem cells (MSCs) has been proposed as a potential cell therapy for cartilage defects. This cell therapy relies on the survival of the implanted MSCs. However, the arduous local environment may limit cell viability after implantation, which would restrict the cells' regenerative capacity. Thus, it is necessary to reinforce the implanted cells against the unfavourable microenvironment in order to improve the efficacy of cell therapy. We examined whether the transduction of an anti‐apoptotic protein, Bcl‐xL, into MSCs could prevent cell death and improve the implantation efficiency of MSCs in a rabbit model. Our current findings demonstrate that the group treated with Bcl‐xL‐engineered MSCs could improve cartilage healing both morphologically and histologically when compared with the controls. These results suggest that intra‐articular injection of Bcl‐xL‐engineered MSCs is a potential non‐invasive therapeutic method for effectively treating cartilage defects of the knee. Copyright © 2009 John Wiley & Sons, Ltd. 相似文献
13.
背景:如何更好地以组织工程学方法修复关节软骨缺损并达到良好的远期疗效目前尚无公识。目的:创新性地在膝关节腔内培养兔骨髓间充质干细胞复合同种异体脱钙骨的组织工程软骨。方法:采用全骨髓贴壁筛选法分离培养兔骨髓间充质干细胞,DMEM/F12完全培养基培养,成软骨诱导条件培养基诱导分化。取同种异体兔的髂骨和椎体骨制作成脱钙骨支架,诱导后的骨髓间充质干细胞种植于脱钙骨支架上,培养1d后将细胞支架复合物用筋膜包裹置于兔左膝关节腔内培养,单纯脱钙骨支架筋膜包裹置入右膝关节腔。于培养第4,8,12周分别取材,行大体观察并制成石蜡切片,采用苏木精伊红染色、甲苯胺蓝染色,Ⅱ型胶原免疫组化染色方法进行组织学观察。结果与结论:培养4,8周,细胞一支架组标本Ⅱ型胶原免疫组化的平均吸光度值(A)分别为0.263±0.031,0.340±0.052,单纯支架组标本分别为0.147±0.027,0.165±0.030,两组比较差异有显著性意义(P〈0.05);培养12周细胞支架组标本Ⅱ型胶原免疫组化A值平均为0.362±0.037,标本类似正常软骨外观,Ⅱ型胶原免疫组化反应呈阳性;而单纯支架组脱钙骨支架降解。培养12周细胞一支架组苏木精一伊红染色结果显示细胞数量多,脱钙骨支架基本被吸收;而甲苯胺蓝染色结果显示有被染成紫红色的异染性基质形成。结果提示兔骨髓间充质干细胞复合同种异体脱钙骨可在兔膝关节腔内培养出组织工程软骨。 相似文献
14.
A humanised tissue‐engineered bone model allows species‐specific breast cancer‐related bone metastasis in vivo
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VMC Quent AV Taubenberger JC Reichert LC Martine JA Clements DW Hutmacher D Loessner 《Journal of tissue engineering and regenerative medicine》2018,12(2):494-504
Bone metastases frequently occur in the advanced stages of breast cancer. At this stage, the disease is deemed incurable. To date, the mechanisms of breast cancer‐related metastasis to bone are poorly understood. This may be attributed to the lack of appropriate animal models to investigate the complex cancer cell–bone interactions. In this study, two established tissue‐engineered bone constructs (TEBCs) were applied to a breast cancer‐related metastasis model. A cylindrical medical‐grade polycaprolactone‐tricalcium phosphate scaffold produced by fused deposition modelling (scaffold 1) was compared with a tubular calcium phosphate‐coated polycaprolactone scaffold fabricated by solution electrospinning (scaffold 2) for their potential to generate ectopic humanised bone in NOD/SCID mice. While scaffold 1 was found not suitable to generate a sufficient amount of ectopic bone tissue due to poor ectopic integration, scaffold 2 showed excellent integration into the host tissue, leading to bone formation. To mimic breast cancer cell colonisation to the bone, MDA‐MB‐231, SUM1315, and MDA‐MB‐231BO breast cancer cells were cultured in polyethylene glycol‐based hydrogels and implanted adjacent to the TEBCs. Histological analysis indicated that the breast cancer cells induced an osteoclastic reaction in the TEBCs, demonstrating analogies to breast cancer‐related bone metastasis seen in patients. 相似文献
15.
In vitro fabrication of autologous living tissue‐engineered vascular grafts based on prenatally harvested ovine amniotic fluid‐derived stem cells
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Benedikt Weber Debora Kehl Ulrich Bleul Luc Behr Sébastien Sammut Laura Frese Agnieszka Ksiazek Josef Achermann Gerald Stranzinger Jérôme Robert Bart Sanders Michele Sidler Chad E. Brokopp Steven T. Proulx Thomas Frauenfelder Roman Schoenauer Maximilian Y. Emmert Volkmar Falk Simon P. Hoerstrup 《Journal of tissue engineering and regenerative medicine》2016,10(1):52-70
Amniotic fluid cells (AFCs) have been proposed as a valuable source for tissue engineering and regenerative medicine. However, before clinical implementation, rigorous evaluation of this cell source in clinically relevant animal models accepted by regulatory authorities is indispensable. Today, the ovine model represents one of the most accepted preclinical animal models, in particular for cardiovascular applications. Here, we investigate the isolation and use of autologous ovine AFCs as cell source for cardiovascular tissue engineering applications. Fetal fluids were aspirated in vivo from pregnant ewes (n = 9) and from explanted uteri post mortem at different gestational ages (n = 91). Amniotic non‐allantoic fluid nature was evaluated biochemically and in vivo samples were compared with post mortem reference samples. Isolated cells revealed an immunohistochemical phenotype similar to ovine bone marrow‐derived mesenchymal stem cells (MSCs) and showed expression of stem cell factors described for embryonic stem cells, such as NANOG and STAT‐3. Isolated ovine amniotic fluid‐derived MSCs were screened for numeric chromosomal aberrations and successfully differentiated into several mesodermal phenotypes. Myofibroblastic ovine AFC lineages were then successfully used for the in vitro fabrication of small‐ and large‐diameter tissue‐engineered vascular grafts (n = 10) and cardiovascular patches (n = 34), laying the foundation for the use of this relevant pre‐clinical in vivo assessment model for future amniotic fluid cell‐based therapeutic applications. Copyright © 2013 John Wiley & Sons, Ltd. 相似文献
16.
Axially vascularized tissue‐engineered bone constructs retain their in vivo angiogenic and osteogenic capacity after high‐dose irradiation
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Ahmad Eweida Oliver Frisch Frank A. Giordano Jens Fleckenstein Frederik Wenz Marc A. Brockmann Matthias Schulte Volker J. Schmidt Ulrich Kneser Leila Harhaus 《Journal of tissue engineering and regenerative medicine》2018,12(2):e657-e668
In order to introduce bone tissue engineering to the field of oncological reconstruction, we are investigating for the first time the effect of various doses of ionizing irradiation on axially vascularized bone constructs. Synthetic bone constructs were created and implanted in 32 Lewis rats. Each construct was axially vascularized through an arteriovenous loop made by direct anastomosis of the saphenous vessels. After 2 weeks, the animals received ionizing irradiation of 9 Gy, 12 Gy and 15 Gy, and were accordingly classified to groups I, II and III, respectively. Group IV was not irradiated and acted as a control. Tissue generation, vascularity, cellular proliferation and apoptosis were investigated either 2 or 5 weeks after irradiation through micro‐computed tomography, histomorphometry and real‐time polymerase chain reaction (PCR). At 2 weeks after irradiation, tissue generation and central vascularity were significantly lower and apoptosis was significantly higher in groups II and III than group IV, but without signs of necrosis. Cellular proliferation was significantly lower in groups I and II. After 5 weeks, the irradiated groups showed improvement in all parameters in relation to the control group, indicating a retained capacity for angiogenesis after irradiation. PCR results confirmed the expression of osteogenesis‐related genes in all irradiated groups. Dense collagen was detected 5 weeks after irradiation, and one construct showed discrete islands of bone indicating a retained osteogenic capacity after irradiation. This demonstrates for the first time that axial vascularization was capable of supporting a synthetic bone construct after a high dose of irradiation that is comparable to adjuvant radiotherapy. Copyright © 2016 John Wiley & Sons, Ltd. 相似文献
17.
背景:微骨折术方法简单,操作方便,是治疗关节软骨缺损有效的方法之一,但仍然存在再生软骨为纤维软骨、再生软骨退化等问题。现在学者们主要致力于使用各种方法改良微骨折修复软骨缺损的效果。 目的:探索微骨折处理软骨缺损区域后植入自体骨髓间充质干细胞外基质支架治疗猪膝关节软骨缺损的疗效。 方法:分离并原代培养猪骨髓间充质干细胞,收集其分泌的细胞外基质膜,采用交联、冻干技术将收集的基质膜制备成三维多孔支架。选取小型成年猪,制备双膝股骨髁、股骨滑车部全层软骨缺损模型,深2 mm,直径6 mm;采用自体左右对照模式,右膝作为对照组,使用单纯微骨折治疗软骨缺损,左膝作为实验组,采用微骨折处理软骨缺损区域后,植入预先制备的支架。术后6个月使用番红固绿染色、Masson 染色等评价软骨再生情况,使用Wakitani评分整体评估再生软骨,并测定再生组织糖胺聚糖、DNA含量。 结果与结论:术后6个月,实验组股骨滑车和股骨髁处均可见软骨修复,表面光滑,对照组股骨滑车修复组织表面较平整,股骨髁未见明显修复。实验组股骨滑车和股骨髁再生软骨经番红固绿染色、Masson染色均显示软骨层基质含量丰富,软骨下骨骨小梁密集,对照组软骨层染色不明显,软骨下骨修复欠佳。实验组Wakitani评分、糖胺聚糖含量高于对照组,DNA含量低于对照组(P&lt;0.05)。结果可见微骨折结合自体骨髓间充质干细胞外基质支架修复软骨效果良好,股骨滑车和股骨髁治疗效果无显著差异。 相似文献
18.
Recombinant human collagen III gel for transplantation of autologous skin cells in porcine full‐thickness wounds
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Sari Suomela Mika Hukkanen Antti Siltanen Ari Harjula Jyrki Vuola Esko Kankuri 《Journal of tissue engineering and regenerative medicine》2015,9(12):1386-1393
Complex skin wounds, such as chronic ulcers and deep burns, require lengthy treatments and cause extensive burdens on healthcare and the economy. Use of biomaterials and cell transplantation may improve traditional treatments and promote the healing of difficult‐to‐treat wounds. In this study, we investigated the use of recombinant human collagen III (rhCol‐III) gel as a delivery vehicle for cultured autologous skin cells (keratinocytes only or keratinocyte–fibroblast mixtures). We examined its effect on the healing of full‐thickness wounds in a porcine wound‐healing model. Two Landrace pigs were used for the study. Fourteen deep dermal wounds were created on the back of each pig with an 8 mm biopsy punch. Syringes containing acellular rhCol‐III gel (n = 8) or rhCol‐III gel with autologous keratinocytes (n = 8) or rhCol‐III gel with autologous keratinocytes and fibroblasts (n = 8) were applied into wounds. Untreated wounds were used as controls for the treatment groups (n = 4). We used rhCol‐III gel to manufacture a cell‐delivery syringe containing autologous skin cells. In a full‐thickness wound‐healing model, we observed that rhCol‐III gel enhances early granulation tissue formation. Interestingly, we found cell type‐dependent differences in the stability of rhCol‐III in vivo. Fibroblast‐containing gel was effectively removed from the wound, whereas gels without cells or with keratinocytes only remained intact. Our results demonstrate that the properties of rhCol‐III gel for skin cell transplantation can be significantly altered in a cell type‐dependent manner. Copyright © 2013 John Wiley & Sons, Ltd. 相似文献
19.
Matrilin‐3 codelivery with adipose‐derived mesenchymal stem cells promotes articular cartilage regeneration in a rat osteochondral defect model
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Manjunatha S. Muttigi Byoung Ju Kim Bogyu Choi Arai Yoshie Hemant Kumar Inbo Han Hansoo Park Soo‐Hong Lee 《Journal of tissue engineering and regenerative medicine》2018,12(3):667-675
Matrilin‐3 is an essential extracellular matrix component present only in cartilaginous tissues. Matrilin‐3 exerts chondroprotective effects by regulating an anti‐inflammatory function and extracellular matrix components. We hypothesized that the codelivery of matrilin‐3 with infrapatellar adipose‐tissue‐derived mesenchymal stem cells (Ad‐MSCs) may enhance articular cartilage regeneration. Matrilin‐3 treatment of Ad‐MSCs in serum‐free media induced collagen II and aggrecan expression, and matrilin‐3 in chondrogenic media also enhanced in vitro chondrogenic differentiation. Next, the in vivo effect of matrilin‐3 codelivery with Ad‐MSCs on cartilage regeneration was assessed in an osteochondral defect model in Sprague Dawley rats: Ad‐MSCs and hyaluronic acid were implanted at the defect site with or without matrilin‐3 (140, 280, and 700 ng). Safranin O staining revealed that matrilin‐3 (140 and 280 ng) treatment significantly improved cartilage regeneration and glycosaminoglycan accumulation. In the animals treated with 140‐ng matrilin‐3, in particular, the defect site exhibited complete integration with surrounding tissue and a smooth glistening surface. The International Cartilage Repair Society macroscopic and O'Driscoll microscopic scores for regenerated cartilage were furthermore shown to be considerably higher for this group (matrilin‐3; 140 ng) compared with the other groups. Furthermore, the defects treated with 140‐ng matrilin‐3 revealed significant hyaline‐like cartilage regeneration in the osteochondral defect model; in contrast, the defects treated with 700‐ng matrilin‐3 exhibited drastically reduced cartilage regeneration with mixed hyaline–fibrocartilage morphology. Codelivery of matrilin‐3 with Ad‐MSCs significantly influenced articular cartilage regeneration, supporting the potential use of this tissue‐specific protein for a cartilage‐targeted stem cell therapy. 相似文献
20.
The effects of different vascular carrier patterns on the angiogenesis and osteogenesis of BMSC–TCP‐based tissue‐engineered bone in beagle dogs
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Ning Kang Jingguo Wu Congmin Gu Jianhai Bi Tao Lv Fangnan Xie Jiewei Hu Xia Liu Yilin Cao Ran Xiao 《Journal of tissue engineering and regenerative medicine》2017,11(2):542-552
Bone repair using tissue‐engineered bone (TEB) in a large defect or accompanied by a poor recipient vascular bed is a long‐standing challenge. Surgical vascular carrier patterns of vascular bundle (VB) and arteriovenous loop (AV loop) have been shown to improve the vascularization and repair capacity of TEB. However, the effects of these different vascular carrier patterns on angiogenesis and osteogenesis in TEB have never been evaluated. Here, TEB was constructed with bone marrow mesenchymal stem cells (BMSCs) and β‐TCP and prevascularized by the VB or AV loop technique in beagle dogs. The vascularization and bone formation in TEB were quantitatively compared using Microfil perfusion, histological examination and CT and micro‐CT analyses. The distribution and constitution of the neovasculature were analysed to determine the underlying mechanism of angiogenesis. The results showed that prevascularized TEB generated bone tissue faster and more homogeneously than untreated TEB. The VB technique was found to strike a better balance between bone regeneration and β‐TCP scaffold degradation than the AV loop strategy, which resulted in more vascularization but less bone yield, due to faster degradation of the β‐TCP scaffold. This study indicates that a suitable triangular relationship, composed of bone regeneration, scaffold degradation and vasculature, is critical to TEB construction. Copyright © 2015 John Wiley & Sons, Ltd. 相似文献