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1.
《Expert opinion on biological therapy》2013,13(11):1601-1622
Introduction: Tissue engineering is a growing area of biomedical research, holding great promise for a broad range of potential applications in the field of regenerative medicine. In recent decades, multiple tissue engineering strategies have been adopted to mimic and improve specific biological functions of tissues and organs, including biomimetic materials, drug-releasing scaffolds, stem cells, and dynamic culture systems. MicroRNAs (miRNAs), noncoding small RNAs that negatively regulate the expression of downstream target mRNAs, are considered a novel class of molecular targets and therapeutics that may play an important role in tissue engineering.Areas covered: Herein, we highlight the latest achievements in regenerative medicine, focusing on the role of miRNAs as key modulators of gene expression, stem cell self-renewal, proliferation and differentiation, and eventually in driving cell fate decisions. Finally, we will discuss the contribution of miRNAs in regulating the rearrangement of the tissue microenvironment and angiogenesis, and the range of strategies for miRNA delivery into target cells and tissues.Expert opinion: Manipulation of miRNAs is an alternative approach and an attractive strategy for controlling several aspects of tissue engineering, although some issues concerning their in vivo effects and optimal delivery methods still remain uncovered. 相似文献
2.
Jun Liu Janneke Hilderink Tom A.M. Groothuis Cees Otto Clemens A. van Blitterswijk Jan de Boer 《Journal of tissue engineering and regenerative medicine》2015,9(8):952-960
Limited nutrient diffusion in three‐dimensional (3D) constructs is a major concern in tissue engineering. Therefore, monitoring nutrient availability and diffusion within a scaffold is an important asset. Since nutrients come in various forms, we have investigated the diffusion of the oxygen, luciferin and dextran molecules within tissue‐engineered constructs using optical imaging technologies. First, oxygen availability and diffusion were investigated, using transgenic cell lines in which a hypoxia‐responsive element drives expression of the green fluorescent protein gene. Using confocal imaging, we observed oxygen limitation, starting at around 200 µm from the periphery in the context of agarose gel with 1 million CHO cells. Diffusion of luciferin was monitored real‐time in agarose gels using a cell line in which the luciferase gene was driven by a constitutively active CMV promoter. Gel concentration affected the diffusion rate of luciferin. Furthermore, we assessed the diffusion rates of fluorescent dextran molecules of different molecular weights in biomaterials by fluorescence recovery after photobleaching (FRAP) and observed that diffusion depended on both molecular size and gel concentration. In conclusion, we have validated a set of efficient tools to investigate molecular diffusion of a range of molecules and to optimize biomaterials design in order to improve nutrient delivery. Copyright © 2013 John Wiley & Sons, Ltd. 相似文献
3.
Review of vascularised bone tissue‐engineering strategies with a focus on co‐culture systems
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Yuchun Liu Jerry K Y Chan Swee‐Hin Teoh 《Journal of tissue engineering and regenerative medicine》2015,9(2):85-105
Poor angiogenesis within tissue‐engineered grafts has been identified as a main challenge limiting the clinical introduction of bone tissue‐engineering (BTE) approaches for the repair of large bone defects. Thick BTE grafts often exhibit poor cellular viability particularly at the core, leading to graft failure and lack of integration with host tissues. Various BTE approaches have been explored for improving vascularisation in tissue‐engineered constructs and are briefly discussed in this review. Recent investigations relating to co‐culture systems of endothelial and osteoblast‐like cells have shown evidence of BTE efficacy in increasing vascularization in thick constructs. This review provides an overview of key concepts related to bone formation and then focuses on the current state of engineered vascularized co‐culture systems using bone repair as a model. It will also address key questions regarding the generation of clinically relevant vascularized bone constructs as well as potential directions and considerations for research with the objective of pursuing engineered co‐culture systems in other disciplines of vascularized regenerative medicine. The final objective is to generate serious and functional long‐lasting vessels for sustainable angiogenesis that will enable enhanced cellular survival within thick voluminous bone grafts, thereby aiding in bone formation and remodelling in the long term. However, more evidence about the quality of blood vessels formed and its associated functional improvement in bone formation as well as a mechanistic understanding of their interactions are necessary for designing better therapeutic strategies for translation to clinical settings. Copyright © 2012 John Wiley & Sons, Ltd. 相似文献
4.
Crina E. Tiron Monica Hellesøy David R. Micklem Jacques E. Nör Lars A. Akslen James B. Lorens 《Journal of tissue engineering and regenerative medicine》2011,5(4):e52-e62
The success of tissue engineering depends on the rapid and efficient formation of a functional blood vasculature. Adult blood vessels comprise endothelial cells and perivascular mural cells that assemble into patent tubules ensheathed by a basement membrane during angiogenesis. Using individual vessel components, we characterized intra‐scaffold microvessel self‐assembly efficiency in a physiological in vivo tissue engineering implant context. Primary human microvascular endothelial and vascular smooth muscle cells were seeded at different ratios in poly‐L ‐lactic acid (PLLA) scaffolds enriched with basement membrane proteins (Matrigel) and implanted subcutaneously into immunocompromised mice. Temporal intra‐scaffold microvessel formation, anastomosis and perfusion were monitored by immunohistochemical, flow cytometric and in vivo multiphoton fluorescence microscopy analysis. Vascularization in the tissue‐engineering context was strongly enhanced in implants seeded with a complete complement of blood vessel components: human microvascular endothelial and vascular smooth muscle cells in vivo assembled a patent microvasculature within Matrigel‐enriched PLLA scaffolds that anastomosed with the host circulation during the first week of implantation. Multiphoton fluorescence angiographic analysis of the intra‐scaffold microcirculation showed a uniform, branched microvascular network. 3D image reconstruction analysis of human pulmonary artery smooth muscle cell (hPASMC) distribution within vascularized implants was non‐random and displayed a preferential perivascular localization. Hence, efficient microvessel self‐assembly, anastomosis and establishment of a functional microvasculture in the native hypoxic in vivo tissue engineering context is promoted by providing a complete set of vascular components. Copyright © 2010 John Wiley & Sons, Ltd. 相似文献
5.
Design and characterization of a biodegradable double‐layer scaffold aimed at periodontal tissue‐engineering applications
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João F. Requicha Carlos A. Viegas Shantesh Hede Isabel B. Leonor Rui L. Reis Manuela E. Gomes 《Journal of tissue engineering and regenerative medicine》2016,10(5):392-403
The inefficacy of the currently used therapies in achieving the regeneration ad integrum of the periodontium stimulates the search for alternative approaches, such as tissue‐engineering strategies. Therefore, the core objective of this study was to develop a biodegradable double‐layer scaffold for periodontal tissue engineering. The design philosophy was based on a double‐layered construct obtained from a blend of starch and poly‐ε‐caprolactone (30:70 wt%; SPCL). A SPCL fibre mesh functionalized with silanol groups to promote osteogenesis was combined with a SPCL solvent casting membrane aiming at acting as a barrier against the migration of gingival epithelium into the periodontal defect. Each layer of the double‐layer scaffolds was characterized in terms of morphology, surface chemical composition, degradation behaviour and mechanical properties. Moreover, the behaviour of seeded/cultured canine adipose‐derived stem cells (cASCs) was assessed. In general, the developed double‐layered scaffolds demonstrated adequate degradation and mechanical behaviour for the target application. Furthermore, the biological assays revealed that both layers of the scaffold allow adhesion and proliferation of the seeded undifferentiated cASCs, and the incorporation of silanol groups into the fibre‐mesh layer enhance the expression of a typical osteogenic marker. This study allowed an innovative construct to be developed, combining a three‐dimensional (3D) scaffold with osteoconductive properties and with potential to assist periodontal regeneration, carrying new possible solutions to current clinical needs. Copyright © 2013 John Wiley & Sons, Ltd. 相似文献
6.
Emilia Krajewska Claire Lewis Carolyn Staton Alice MacGowan Sheila MacNeil 《Journal of tissue engineering and regenerative medicine》2011,5(5):363-374
We have previously shown that putrescine induces a psoriatic phenotype in tissue‐engineered skin. The initial aim of this study was to further develop this in vitro model by introducing endothelial cells to mimic the increased vascularization found in psoriasis. Human keratinocytes and fibroblasts, which did not express CD34 or CD31 in 2D culture, were added to de‐epidermised acellular human dermis and cultured for 4 weeks. For induction of a psoriatic phenotype, putrescine was added during this period. We report that after 4 weeks of culture, and particularly when exposed to putrescine, this model showed expression of vertically organised clusters of CD31 positive cells in the dermis in the absence of any exogenous endothelial cells. Further investigation in 2D cell cultures showed an indirect effect of putrescine on normal keratinocytes causing them to produce soluble factors that increased expression of CD133, CD34 and CD31 in cultured human dermal fibroblasts, previously negative for these antigens. This study reports a new and improved model of psoriasis for in vitro studies and offers a new insight into early stage neovascularization, which is of relevance not only to psoriasis, but to tissue engineering and wound healing in general. Copyright © 2010 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.
Design and validation of a dynamic cell‐culture system for bone biology research and exogenous tissue‐engineering applications
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Alexander C. Allori Edward H. Davidson Derek D. Reformat Alexander M. Sailon James Freeman Adam Vaughan David Wootton Elizabeth Clark John L. Ricci Stephen M. Warren 《Journal of tissue engineering and regenerative medicine》2016,10(10):E327-E336
Bone lacunocanalicular fluid flow ensures chemotransportation and provides a mechanical stimulus to cells. Traditional static cell‐culture methods are ill‐suited to study the intricacies of bone biology because they ignore the three‐dimensionality of meaningful cellular networks and the lacunocanalicular system; furthermore, reliance on diffusion alone for nutrient supply and waste product removal effectively limits scaffolds to 2–3 mm thickness. In this project, a flow‐perfusion system was custom‐designed to overcome these limitations: eight adaptable chambers housed cylindrical cell‐seeded scaffolds measuring 12 or 24 mm in diameter and 1–10 mm in thickness. The porous scaffolds were manufactured using a three‐dimensional (3D) periodic microprinting process and were composed of hydroxyapatite/tricalcium phosphate with variable thicknesses, strut sizes, pore sizes and structural configurations. A multi‐channel peristaltic pump drew medium from parallel reservoirs and perfused it through each scaffold at a programmable rate. Hermetically sealed valves permitted sampling or replacement of medium. A gas‐permeable membrane allowed for gas exchange. Tubing was selected to withstand continuous perfusion for > 2 months without leakage. Computational modelling was performed to assess the adequacy of oxygen supply and the range of fluid shear stress in the bioreactor–scaffold system, using 12 × 6 mm scaffolds, and these models suggested scaffold design modifications that improved oxygen delivery while enhancing physiological shear stress. This system may prove useful in studying complex 3D bone biology and in developing strategies for engineering thick 3D bone constructs. Copyright © 2013 John Wiley & Sons, Ltd. 相似文献
9.
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
10.
Motility imaging via optical coherence phase microscopy enables label‐free monitoring of tissue growth and viability in 3D tissue‐engineering scaffolds
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Christina Holmes Maryam Tabrizian Pierre O. Bagnaninchi 《Journal of tissue engineering and regenerative medicine》2015,9(5):641-645
As the field of tissue engineering continues to progress, there is a deep need for non‐invasive, label‐free imaging technologies that can monitor tissue growth and health within thick three‐dimensional (3D) constructs. Amongst the many imaging modalities under investigation, optical coherence tomography (OCT) has emerged as a promising tool, enabling non‐destructive in situ characterization of scaffolds and engineered tissues. However, the lack of optical contrast between cells and scaffold materials using this technique remains a challenge. In this communication, we show that mapping the optical phase fluctuations resulting from cellular viability and motility allows for the distinction of live cells from their surrounding scaffold environment. Motility imaging was performed via a common‐path optical coherence phase microscope (OCPM), an OCT modality that has been shown to be sensitive to nanometer‐level fluctuations. More specifically, we examined the development of human adipose‐derived stem cells and/or murine pre‐osteoblasts within two distinct scaffold systems, commercially available alginate sponges and custom‐microfabricated poly(d , l ‐lactic‐co‐glycolic acid) fibrous scaffolds. Cellular motility is demonstrated as an endogenous source of contrast for OCPM, enabling real‐time, label‐free monitoring of 3D engineered tissue development. Copyright © 2013 John Wiley & Sons, Ltd. 相似文献
11.
Matthias W. Laschke Philipp Karschnia Claudia Scheuer Alexander Heß Wolfgang Metzger Michael D. Menger 《Journal of tissue engineering and regenerative medicine》2018,12(4):1020-1030
Adipose tissue‐derived microvascular fragments (ad‐MVF) are effective vascularization units for tissue engineering. They rapidly reassemble into new microvascular networks after seeding on scaffolds and subsequent in vivo implantation. Herein, we analyzed whether the vascularization capacity of ad‐MVF is affected by cryopreservation. Ad‐MVF were isolated from the epididymal fat pads of C57BL/6 mice and cryopreserved for 7 days to compare their morphology, viability, cellular composition, and protein expression with freshly isolated control ad‐MVF. Moreover, cryopreserved and control ad‐MVF from green fluorescent protein (GFP)+ donor mice were seeded on collagen‐glycosaminoglycan matrices (Integra®), which were implanted into dorsal skinfold chambers of GFP? recipient animals to study their vascularization and incorporation using intravital fluorescence microscopy, histology, and immunohistochemistry. Cryopreservation of ad‐MVF did not affect vessel morphology and cellular composition. However, cryopreservation was associated with an increased rate of necrotic cells and a significantly reduced number of transplantable ad‐MVF. This was compensated by a higher angiogenic activity of the remaining ad‐MVF, as indicated by significantly elevated expression levels of pro‐angiogenic factors when compared to controls. Accordingly, cryopreserved and control ad‐MVF induced a comparable vascularization and incorporation of implanted Integra® without differences in microvascular network formation, maturation, and remodeling. Enhanced angiogenic sprouting even resulted in a higher fraction of GFP+ microvessels within the implants of the cryopreservation group. These findings indicate that cryopreservation of ad‐MVF is feasible and, thus, offers the exciting opportunity to build up stocks of readily available vascularization units for future tissue engineering applications. 相似文献
12.
Glottic regeneration with a tissue‐engineering technique,using acellular extracellular matrix scaffold in a canine model
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Morimasa Kitamura Shigeru Hirano Shin‐ichi Kanemaru Yoshiharu Kitani Satoshi Ohno Tsuyoshi Kojima Tatsuo Nakamura Juichi Ito Clark A. Rosen Thomas W. Gilbert 《Journal of tissue engineering and regenerative medicine》2016,10(10):825-832
Acellular extracellular matrix scaffold derived from porcine urinary bladder (UBM) is decellularized material that has shown success for constructive remodelling of various tissues and organs. The regenerative effects of UBM were reported for the tympanic membrane, oesophagus, trachea, larynx, pleura and pericardium in animal studies, with promising results. The aim of this study was to investigate the regenerative effects of UBM on hemilarynx, using a canine model. A left partial hemilaryngectomy was performed and the surgical defects were reconstructed by insertion of UBM scaffold. Although local infection was observed in one dog in 1 week after implantation of the scaffold, all dogs showed good re‐epithelialization with minimum complication in 1 month. The effect of regeneration of the larynx was evaluated 6 months after the operation. The excised larynx experiments were performed to measure phonation threshold pressure (PTP), normalized mucosal wave amplitude (NMWA) and normalized glottal gap (NGG). The results of the measurements showed that PTP was normal or near normal in two cases and NMWA was within normal range in three cases, although there were individual variations. Histological examination was completed to evaluate structural changes in the scaffold with the appearance of the new cartilaginous structure. However, the regenerated vocal fold mucosa was mostly scarred. The UBM scaffold has shown to be biocompatible, biodegradable and useful for tissue regeneration of the hemilarynx, with possible restoration of function of the vocal fold. The vocal fold mucosa was scarred, which is the next challenge to be addressed. Copyright © 2014 John Wiley & Sons, Ltd. 相似文献
13.
Ilyas Inci Harald Kirsebom Igor Yu Galaev Bo Mattiasson Erhan Piskin 《Journal of tissue engineering and regenerative medicine》2013,7(7):584-588
Gelatin‐based cryogels were prepared by using a novel crosslinker, oxidized dextran, which was synthesized and used in the study. The cryogels were also loaded with freshly formed hydroxyapatite (HA) particles. These cryogels are opaque, spongy and highly elastic and have a pore structure with large interconnected pores. They swell about 500% in aqueous media and within a few minutes reach their final swollen forms. The elastic moduli of HA‐containing cryogels were 18.5 ± 3.0 kPa, which is suitable for non‐load‐bearing bone tissue‐engineering (TE) applications, especially for the craniofacial area. Copyright © 2012 John Wiley & Sons, Ltd. 相似文献
14.
Mamoni Dash Sangram K. Samal Timothy E. L. Douglas David Schaubroeck Sander C. Leeuwenburgh Pascal Van Der Voort Heidi A. Declercq Peter Dubruel 《Journal of tissue engineering and regenerative medicine》2017,11(5):1500-1513
Porous biodegradable scaffolds represent promising candidates for tissue‐engineering applications because of their capability to be preseeded with cells. We report an uncrosslinked chitosan scaffold designed with the aim of inducing and supporting enzyme‐mediated formation of apatite minerals in the absence of osteogenic growth factors. To realize this, natural enzyme alkaline phosphatase (ALP) was incorporated into uncrosslinked chitosan scaffolds. The uncrosslinked chitosan makes available amine and alcohol functionalities to enhance the biomineralization process. The physicochemical findings revealed homogeneous mineralization, with the phase structure of the formed minerals resembling that of apatite at low mineral concentrations, and similar to dicalcium phosphate dihydrate (DCPD) with increasing ALP content. The MC3T3 cell activity clearly showed that the mineralization of the chitosan scaffolds was effective in improving cellular adhesion, proliferation and colonization. Copyright © 2015 John Wiley & Sons, Ltd. 相似文献
15.
Platelet‐rich fibrin‐based matrices to improve angiogenesis in an in vitro co‐culture model for bone tissue engineering
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Eva Dohle Karima El Bagdadi Robert Sader Joseph Choukroun C. James Kirkpatrick Shahram Ghanaati 《Journal of tissue engineering and regenerative medicine》2018,12(3):598-610
In the context of prevascularization strategies for tissue‐engineering purposes, co‐culture systems consisting of outgrowth endothelial cells (OECs) and primary osteoblasts (pOBs) have been established as a promising in vitro tool to study regeneration mechanisms and to identify factors that might positively influence repair processes such as wound healing or angiogenesis. The development of autologous injectable platelet‐rich fibrin (PRF), which can be generated from peripheral blood in a minimal invasive procedure, fulfils several requirements for clinically applicable cell‐based tissue‐engineering strategies. During this study, the established co‐culture system of OECs and pOBs was mixed with injectable PRF and was cultivated in vitro for 24 h or 7 days. The aim of this study was to analyse whether PRF might have a positive effect on wound healing processes and angiogenic activation of OECs in the co‐culture with regard to proinflammatory factors, adhesion molecules and proangiogenic growth factor expression. Histological cell detection revealed the formation of lumina and microvessel‐like structures in the PRF/co‐culture complexes after 7 days of complex cultivation. Interestingly, the angiogenic activation of OECs was accompanied by an upregulation of wound healing‐associated factors, as well as by a higher expression of the proangiogenic factor vascular endothelial growth factor, which was evaluated both on the mRNA level as well as on the protein level. Thus, PRF might positively influence wound healing processes, in particular angiogenesis, in the in vitro co‐culture, making autologous PRF‐based matrices a beneficial therapeutic tool for tissue‐engineering purposes by simply profiting from the PRF, which contains blood plasma, platelets and leukocytes. 相似文献
16.
Sajedeh Khorshidi Atefeh Solouk Hamid Mirzadeh Saeedeh Mazinani Jose M. Lagaron Shahriar Sharifi Seeram Ramakrishna 《Journal of tissue engineering and regenerative medicine》2016,10(9):715-738
Tissue engineering holds great promise to develop functional constructs resembling the structural organization of native tissues to improve or replace biological functions, with the ultimate goal of avoiding organ transplantation. In tissue engineering, cells are often seeded into artificial structures capable of supporting three‐dimensional (3D) tissue formation. An optimal scaffold for tissue‐engineering applications should mimic the mechanical and functional properties of the extracellular matrix (ECM) of those tissues to be regenerated. Amongst the various scaffolding techniques, electrospinning is an outstanding one which is capable of producing non‐woven fibrous structures with dimensional constituents similar to those of ECM fibres. In recent years, electrospinning has gained widespread interest as a potential tissue‐engineering scaffolding technique and has been discussed in detail in many studies. So why this review? Apart from their clear advantages and extensive use, electrospun scaffolds encounter some practical limitations, such as scarce cell infiltration and inadequate mechanical strength for load‐bearing applications. A number of solutions have been offered by different research groups to overcome the above‐mentioned limitations. In this review, we provide an overview of the limitations of electrospinning as a tissue‐engineered scaffolding technique, with emphasis on possible resolutions of those issues. Copyright © 2015 John Wiley & Sons, Ltd. 相似文献
17.
Matthias W. Laschke Alexander Heß Claudia Scheuer Philipp Karschnia Michael D. Menger 《Journal of tissue engineering and regenerative medicine》2019,13(2):131-142
Adipose tissue‐derived microvascular fragments (ad‐MVFs) are promising vascularization units for tissue engineering. In this study, we analysed the effects of normothermic (37°C) and subnormothermic (20°C) short‐term cultivation on their viability and network forming capacity. Ad‐MVFs from green fluorescent protein (GFP)+ and GFP? C57BL/6 mice were cultivated for 24 hr at 37°C or 20°C. Freshly isolated, noncultivated ad‐MVFs served as controls. Number, length, viability, proliferation, and angiogenic activity of the ad‐MVFs were assessed by microscopic analysis and proteome profiling. GFP+ ad‐MVFs were seeded onto collagen‐glycosaminoglycan matrices, which were implanted into dorsal skinfold chambers of GFP? mice to analyse their vascularization by means of intravital fluorescence microscopy, histology, and immunohistochemistry. Depending on the temperature, short‐term cultivation of ad‐MVFs markedly changed their expression of multiple proangiogenic and antiangiogenic factors. Moreover, cultivation at 37°C significantly increased the number of apoptotic cells within ad‐MVFs, whereas 20°C preserved the viability of ad‐MVFs and even promoted the proliferation of endothelial and perivascular cells. Accordingly, ad‐MVFs cultivated at 20°C also exhibited an enhanced in vivo vascularization capacity when compared with normothermically cultivated ad‐MVFs and noncultivated controls. This was indicated by an accelerated network formation, an increased microvascular remodelling, and a higher density of GFP+ microvessels within implanted matrices. Thus, if ad‐MVFs require short‐term storage before in vivo application, subnormothermic cultivation should be preferred to normothermic cultivation. 相似文献
18.
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
19.
Leko Lin Anat Perets Yah‐el Har‐el Devika Varma Mengyan Li Philip Lazarovici Dara L. Woerdeman Peter I. Lelkes 《Journal of tissue engineering and regenerative medicine》2013,7(12):994-1008
As a potential alternative to currently available skin substitutes and wound dressings, we explored the use of bioactive scaffolds made of plant‐derived proteins. We hypothesized that ‘green’ materials, derived from renewable and biodegradable natural sources, may confer bioactive properties to enhance wound healing and tissue regeneration. We optimized and characterized fibrous scaffolds electrospun from soy protein isolate (SPI) with addition of 0.05% poly(ethylene oxide) (PEO) dissolved in 1,1,1,3,3,3‐hexafluoro‐2‐propanol, and from corn zein dissolved in glacial acetic acid. Fibrous mats electrospun from either of these plant proteins remained intact without further cross‐linking, possessing a skin‐like pliability. Soy‐derived scaffolds supported the adhesion and proliferation of cultured primary human dermal fibroblasts. Using targeted PCR arrays and qPCR validation, we found similar gene expression profiles of fibroblasts cultured for 2 and 24 h on SPI substrates and on collagen type I at both time points. On both substrates there was a pronounced time‐dependent upregulation of several genes related to ECM deposition remodelling, including MMP‐10, MMP‐1, collagen VII, integrin‐α2 and laminin‐β3, indicating that both plant‐ and animal‐derived materials induce similar responses from the cells after initial adhesion, degrading substrate proteins and depositing extracellular matrix in a ‘normal’ remodelling process. These results suggest that ‘green’ proteins, such as soy and zein, are promising as a platform for organotypic skin equivalent culture, as well as implantable scaffolds for skin regeneration. Future studies will determine specific mechanisms of their interaction with skin cells and their efficacy in wound‐healing applications. Copyright © 2012 John Wiley & Sons, Ltd. 相似文献
20.
Horseradish peroxidase‐catalysed in situ‐forming hydrogels for tissue‐engineering applications
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Jin Woo Bae Jong Hoon Choi Yunki Lee Ki Dong Park 《Journal of tissue engineering and regenerative medicine》2015,9(11):1225-1232
In situ‐forming hydrogels are an attractive class of implantable biomaterials that are used for biomedical applications. These injectable hydrogels are versatile and provide a convenient platform for delivering cells and drugs via minimally invasive surgery. Although several crosslinking methods for preparing in situ forming hydrogels have been developed over the past two decades, most hydrogels are not sufficiently versatile for use in a wide variety of tissue‐engineering applications. In recent years, enzyme‐catalysed crosslinking approaches have been emerged as a new approach for developing in situ‐forming hydrogels. In particular, the horseradish peroxidase (HRP)‐catalysed crosslinking approach has received increasing interest, due to its highly improved and tunable capacity to obtain hydrogels with desirable properties. The HRP‐catalysed crosslinking reaction immediately occurs upon mixing phenol‐rich polymers with HRP and hydrogen peroxide (H2O2) in aqueous media. Based on this unique gel‐forming feature, recent studies have shown that various properties of formed hydrogels, such as gelation time, stiffness and degradation rate, can be easily manipulated by varying the concentrations of HRP and H2O2. In this review, we outline the versatile properties of HRP‐catalysed in situ‐forming hydrogels, with a brief introduction to the crosslinking mechanisms involved. In addition, the recent biomedical applications of HRP‐catalysed in situ‐forming hydrogels for tissue regeneration are described. Copyright © 2014 John Wiley & Sons, Ltd. 相似文献