RESUMO
While neural cell transplantation represents a promising therapy for neurodegenerative diseases, the formation of functional networks of transplanted cells with host neurons constitutes one of the challenging steps. Here, we introduce a magnetic guidance methodology that controls neurite growth signaling via magnetic nanoparticles (MNPs) conjugated with antibodies targeting the deleted in colorectal cancer (DCC) receptor (DCC-MNPs). Activation of the DCC receptors by clusterization and subsequent axonal growth of the induced neuronal (iN) cells was performed in a spatially controlled manner. In addition to the directionality of the magnetically controlled axon projection, axonal growth of the iN cells assisted the formation of functional connections with pre-existing primary neurons. Our results suggest magnetic guidance as a strategy for improving neuronal connectivity by spatially guiding the axonal projections of transplanted neural cells for synaptic interactions with the host tissue.
Assuntos
Anticorpos/química , Axônios/metabolismo , Reprogramação Celular , Receptor DCC/metabolismo , Células-Tronco Pluripotentes Induzidas/metabolismo , Campos Magnéticos , Nanopartículas de Magnetita/química , Receptor DCC/antagonistas & inibidores , Humanos , Células-Tronco Pluripotentes Induzidas/citologia , Neuritos/metabolismoRESUMO
Sound perception via mechano-sensation is a remarkably sensitive and fast transmission process, converting sound as a mechanical input to neural signals in a living organism. Although knowledge of auditory hair cell functions has advanced over the past decades, challenges remain in understanding their biomechanics, partly because of their biophysical complexity and the lack of appropriate probing tools. Most current studies of hair cells have been conducted in a relatively low-frequency range (<1000 Hz); therefore, fast kinetic study of hair cells has been difficult, even though mammalians have sound perception of 20 kHz or higher. Here, we demonstrate that the magnetic force nanoprobe (MFN) has superb spatiotemporal capabilities to mechanically stimulate spatially-targeted individual hair cells with a temporal resolution of up to 9 µs, which is equivalent to approximately 50 kHz; therefore, it is possible to investigate avian hair cell biomechanics at different tonotopic regions of the cochlea covering a full hearing frequency range of 50 to 5000 Hz. We found that the variation of the stimulation frequency and amplitude of hair bundles creates distinct mechanical responsive features along the tonotopic axis, where the kinetics of the hair bundle recovery motion exhibits unique frequency-dependent characteristics: basal, middle, and apical hair bundles can effectively respond at their respective ranges of frequency. We revealed that such recovery kinetics possesses two different time constants that are closely related to the passive and active motilities of hair cells. The use of MFN is critical for the kinetics study of free-standing hair cells in a spatiotemporally distinct tonotopic organization.
Assuntos
Células Ciliadas Auditivas/fisiologia , Nanopartículas de Magnetita/química , Som , Animais , Fenômenos Biomecânicos , Galinhas , Cinética , Campos Magnéticos , Imãs , Tamanho da Partícula , Propriedades de SuperfícieRESUMO
For severe burn injuries, successful medical intervention is accomplished by rapidly and safely providing physical barriers that can cover damaged skin tissues, thereby preventing critical danger of extensive bleeding and infection. Despite availability of a large assortment of wound coverage options, the etiology of wound healing is rather complex leading to significant defects in skin repair. The use of cell-mediated treatment approaches in combination with bioengineered wound coverage constructs may provide the missing tool to improve wound healing outcomes. In this study, we have used an engineered 3D PEGylated fibrin (P-fibrin) gel as a scaffold for adipose derived stem cells (ASCs) delivery into the burn injury model. We were able to confirm the presence of ASCs in the wound site two weeks after the initial injury. Delivery of ASCs-containing gels was associated with improved vascularization of the injured area at early time points accompanied by an increased abundance of mannose receptor expressing cells. Moreover, the application of P-fibrin biomaterial exhibited positive effects on early mononuclear cell recruitment and granulation tissue formation without negatively affecting wound closure kinetics or extent of connective tissue deposition. Collectively, our data support the feasibility of using P-fibrin gels in wound dressing applications requiring controlled delivery of viable cells.
RESUMO
The bioengineering of autologous vascular networks is of great importance in wound healing. Adipose-derived stem cells (ASCs) are of interest due to their ability to differentiate toward various cell types, including vascular. We hypothesized that adult human ASCs embedded in a three-dimensional PEG-fibrin (FPEG) gel have the ability to modulate vascularization of a healing wound. Initial in vitro characterization of ASCs isolated from discarded burn skin samples (dsASCs) and embedded in FPEG gels indicated they could express such pericyte/smooth muscle cell markers as α-smooth muscle actin, platelet-derived growth factor receptor-ß, NG2 proteoglycan, and angiopoietin-1, suggesting that these cells could potentially be involved in a supportive cell role (i.e., pericyte/mural cell) for blood vessels. Using a rat skin excision model, wounds treated with dsASCs-FPEG gels showed earlier collagen deposition and wound remodeling compared to vehicle FPEG treated wounds. Furthermore, the dsASCs-seeded gels increased the number of vessels in the wound per square millimeter by day 16 (~66.7 vs. ~36.9/mm(2)) in these same studies. dsASCs may support this increase in vascularization through their trophic contribution of vascular endothelial growth factor, as determined by in vitro analysis of mRNA and the protein levels. Immunohistochemistry showed that dsASCs were localized to the surrounding regions of large blood-perfused vessels. Human dsASCs may play a supportive role in the formation of vascular structures in the healing wound through direct mechanisms as well as indirect trophic effects. The merging of autologous grafts or bioengineered composites with the host's vasculature is critical, and the use of autologous dsASCs in these procedures may prove to be therapeutic.
Assuntos
Células-Tronco Adultas/citologia , Queimaduras/patologia , Neovascularização Fisiológica/fisiologia , Regeneração/fisiologia , Pele/irrigação sanguínea , Alicerces Teciduais , Cicatrização/fisiologia , Adulto , Animais , Biomarcadores , Queimaduras/cirurgia , Técnicas de Cultura de Células/métodos , Diferenciação Celular , Células Cultivadas , Desbridamento/efeitos adversos , Matriz Extracelular , Fibrinogênio , Géis , Xenoenxertos , Humanos , Masculino , Polietilenoglicóis , Ratos , Ratos Nus , Pele/lesões , Transplante AutólogoRESUMO
Single-walled carbon nanohorns (SWNHs) are new carbonaceous materials. In this paper, we report the first successful preparation of SWNHs encapsulating trimetallic nitride template endohedral metallofullerenes (TNT-EMFs). The resultant materials were functionalized by a high-speed vibration milling method and conjugated with CdSe/ZnS quantum dots (QDs). The successful encapsulation of TNT-EMFs and external functionalization with QDs provide a dual diagnostic platform for in vitro and in vivo biomedical applications of these new carbonaceous materials.
Assuntos
Fulerenos/química , Metais/química , Nanotubos de Carbono , Pontos Quânticos , Técnicas In VitroRESUMO
Parkinson's disease (PD) is a progressive neurodegenerative disorder characterized by mitochondrial dysfunction, Lewy body formation, and loss of dopaminergic neurons. Parkin, an E3 ubiquitin ligase, is thought to inhibit PD progression by removing damaged mitochondria and suppressing the accumulation of α-synuclein and other protein aggregates. The present study describes a protein-based therapy for PD enabled by the development of a cell-permeable Parkin protein (iCP-Parkin) with enhanced solubility and optimized intracellular delivery. iCP-Parkin recovered damaged mitochondria by promoting mitophagy and mitochondrial biogenesis and suppressed toxic accumulations of α-synuclein in cells and animals. Last, iCP-Parkin prevented and reversed declines in tyrosine hydroxylase and dopamine expression concomitant with improved motor function induced by mitochondrial poisons or enforced α-synuclein expression. These results point to common, therapeutically tractable features in PD pathophysiology, and suggest that motor deficits in PD may be reversed, thus providing opportunities for therapeutic intervention after the onset of motor symptoms.
Assuntos
Doença de Parkinson , alfa-Sinucleína , Animais , Neurônios Dopaminérgicos/metabolismo , Mitocôndrias/metabolismo , Doença de Parkinson/metabolismo , Ubiquitina-Proteína Ligases/genética , Ubiquitina-Proteína Ligases/metabolismo , alfa-Sinucleína/genéticaRESUMO
AIM: Progressive ischemia due to peripheral artery disease causes muscle damage and reduced strength of the lower extremities. Autologous cell therapy is an attractive treatment to restore perfusion and improve muscle function. Adipose-derived stem cells (ASCs) have therapeutic potential in tissue repair, including polarizing effects on macrophages (MPs). MATERIALS & METHODS: Co-culture systems of ASCs and MPs were analyzed for gene and protein expression modifications in ASC-conditioned MPs. Co-transplantation of MPs/ASCs in vivo led to improved skeletal muscle regeneration in a mouse model of peripheral artery disease. RESULTS: ASCs/MPs therapy restored muscle function, increased perfusion and reduced inflammatory infiltrate. CONCLUSION: Combined MPs/ASCs cell therapy is a promising approach to restore muscle function and stimulate local angiogenesis in the ischemic limb.
Assuntos
Tecido Adiposo/citologia , Terapia Baseada em Transplante de Células e Tecidos , Isquemia/terapia , Macrófagos/citologia , Músculo Esquelético/citologia , Regeneração/fisiologia , Células-Tronco/citologia , Animais , Células Cultivadas , Técnicas de Cocultura , Modelos Animais de Doenças , Feminino , Humanos , Camundongos , Camundongos Endogâmicos C57BL , Músculo Esquelético/lesões , Neovascularização Fisiológica , Doença Arterial Periférica/terapia , Transplante de Células-TroncoRESUMO
Current biomedical imaging tools have limitations in accurate assessment of the severity of open and deep burn wounds involving excess bleeding and severe tissue damage. Furthermore, sophisticated imaging techniques are needed for advanced therapeutic approaches such as noninvasive monitoring of stem cells seeded and applied in a biomedical 3D scaffold to enhance wound repair. This work introduces a novel application of combined ultrasound (US) and photoacoustic (PA) imaging to assess both burn injury and skin tissue regeneration. Tissue structural damage and bleeding throughout the epidermis and dermis till the subcutaneous skin layer were imaged noninvasively by US/PA imaging. Gold nanoparticle-labeled adipose-derived stem cells (ASCs) within a PEGylated fibrin 3D gel were implanted in a rat model of cutaneous burn injury. ASCs were successfully tracked till 2 weeks and were distinguished from host tissue components (e.g., epidermis, fat, and blood vessels) through spectroscopic PA imaging. The structure and function of blood vessels (vessel density and perfusion) in the wound bed undergoing skin tissue regeneration were monitored both qualitatively and semi-quantitatively by the developed imaging approach. Imaging-based analysis demonstrated ASC localization in the top layer of skin and a higher density of regenerating blood vessels in the treated groups. This was corroborated with histological analysis showing localization of fluorescently labeled ASCs and smooth muscle alpha actin-positive blood vessels. Overall, the US/PA imaging-based strategy coupled with gold nanoparticles has a great potential for stem cell therapies and tissue engineering due to its noninvasiveness, safety, selectivity, and ability to provide long-term monitoring.
Assuntos
Tecido Adiposo/metabolismo , Queimaduras/patologia , Técnicas Fotoacústicas/métodos , Pele/patologia , Células-Tronco/metabolismo , Ultrassonografia/métodos , Cicatrização , Tecido Adiposo/patologia , Animais , Queimaduras/terapia , Ouro , Humanos , Masculino , Nanopartículas Metálicas , Ratos , Ratos Endogâmicos Lew , Pele/metabolismo , Transplante de Células-Tronco , Células-Tronco/patologiaRESUMO
Engineered three-dimensional biomaterials are known to affect the regenerative capacity of stem cells. The extent to which these materials can modify cellular activities is still poorly understood, particularly for adipose-derived stem cells (ASCs). This study evaluates PEGylated fibrin (P-fibrin) gels as an ASC-carrying scaffold for encouraging local angiogenesis by comparing with two commonly used hydrogels (i.e., collagen and fibrin) in the tissue-engineering field. Human ASCs in P-fibrin were compared to cultures in collagen and fibrin under basic growth media without any additional soluble factors. ASCs proliferated similarly in all gel scaffolds but showed significantly elongated morphologies in the P-fibrin gels relative to other gels. P-fibrin elicited higher von Willebrand factor expression in ASCs than either collagen or fibrin while cells in collagen expressed more smooth muscle alpha actin than in other gels. VEGF was secreted more at 7 days in fibrin and P-fibrin than in collagen and several other angiogenic and immunomodulatory cytokines were similarly enhanced. Fibrin-based matrices appear to activate angiogenic signaling in ASCs while P-fibrin matrices are uniquely able to also drive a vessel-like ASC phenotype. Collectively, these results suggest that P-fibrin promotes the angiogenic potential of ASC-based therapeutic applications.
Assuntos
Adipócitos/citologia , Tecido Adiposo/citologia , Técnicas de Cultura de Células , Fibrina/química , Neovascularização Fisiológica , Células-Tronco/citologia , Animais , Diferenciação Celular/fisiologia , Células Cultivadas , Colágeno/química , Colágeno/metabolismo , Meios de Cultivo Condicionados , Citocinas/metabolismo , Fibrinogênio/química , Humanos , Hidrogéis/química , Microscopia de Fluorescência , Morfogênese , Fenótipo , Ratos , Medicina Regenerativa , Engenharia Tecidual/métodos , Fator de von Willebrand/metabolismoRESUMO
Stress conditioning (e.g., thermal, shear, and tensile stress) of bone cells has been shown to enhance healing. However, prior studies have not investigated whether combined stress could synergistically promote bone regeneration. This study explored the impact of combined thermal and tensile stress on the induction of heat shock proteins (HSPs) and bone-related proteins by a murine preosteoblast cell line (MC3T3-E1). Cells were exposed to thermal stress using a water bath (44°C for 4 or 8 minutes) with postheating incubation (37°C for 4 hours) followed by exposure to cyclic strain (equibiaxial 3%, 0.2 Hz, cycle of 10-second tensile stress followed by 10-second rest). Combined thermal stress and tensile stress induced mRNA expression of HSP27 (1.41 relative fold induction (RFI) compared to sham-treated control), HSP70 (5.55 RFI), and osteopontin (1.44 RFI) but suppressed matrix metalloproteinase-9 (0.6 RFI) compared to the control. Combined thermal and tensile stress increased vascular endothelial growth factor (VEGF) secretion into the culture supernatant (1.54-fold increase compared to the control). Therefore, combined thermal and mechanical stress preconditioning can enhance HSP induction and influence protein expression important for bone tissue healing.
Assuntos
Regeneração Óssea/genética , Proteínas de Choque Térmico HSP27/biossíntese , Proteínas de Choque Térmico HSP70/biossíntese , Estresse Mecânico , Animais , Linhagem Celular , Sobrevivência Celular/genética , Regulação da Expressão Gênica , Resposta ao Choque Térmico , Calefação , Camundongos , Osteopontina/biossíntese , RNA Mensageiro/biossíntese , Fator A de Crescimento do Endotélio Vascular/biossínteseRESUMO
Evaluating the regenerative capacity of a tissue-engineered device in a noninvasive and synchronous manner is critical to determining the mechanisms for success in clinical applications. In particular, directly tracking implanted cells in a three-dimensional (3D) scaffold is desirable in that it enables the monitoring of cellular activity in a specific and localized manner. The authors' group has previously demonstrated that the PEGylation of fibrin results in a 3D scaffold that supports morphologic and phenotypic changes in mesenchymal stem cells that may be advantageous in wound healing applications. Recently, the authors have evaluated adipose-derived stem cells (ASCs) as a mesenchymal cell source to regenerate skin and blood vessels due to their potential for proliferation, differentiation, and production of growth factors. However, tracking and monitoring ASCs in a 3D scaffold, such as a PEGylated fibrin gel, have not yet been fully investigated. In the current paper, nanoscale gold spheres (20 nm) as cell tracers for ASCs cultured in a PEGylated fibrin gel were evaluated. An advanced dual-imaging modality combining ultrasound and photoacoustic imaging was utilized to monitor rat ASCs over time. The ASCs took up gold nanotracers and could be detected up to day 16 with high sensitivity using photoacoustic imaging. There were no detrimental effects on ASC morphology, network formation, proliferation, and protein expression/secretion (ie, smooth muscle α-actin, vascular endothelial growth factor, matrix metalloproteinase-2, and matrix metalloproteinase-9) associated with gold nanotracers. Therefore, utilization of gold nanotracers can be an effective strategy to monitor the regenerative process of a stem cell source in a 3D gel for vascular and dermal tissue engineering applications.
Assuntos
Tecido Adiposo/citologia , Rastreamento de Células/métodos , Fibrina/química , Nanopartículas Metálicas/química , Células-Tronco/citologia , Engenharia Tecidual/métodos , Análise de Variância , Animais , Fibrina/metabolismo , Ouro/química , Ouro/farmacocinética , Masculino , Metaloproteinase 2 da Matriz/metabolismo , Metaloproteinase 9 da Matriz/metabolismo , Técnicas Fotoacústicas , Polietilenoglicóis/química , Polietilenoglicóis/metabolismo , Ratos , Ratos Endogâmicos Lew , Células-Tronco/química , Células-Tronco/metabolismo , Fator A de Crescimento do Endotélio Vascular/metabolismoRESUMO
Smooth muscle cells (SMC) are critical in stabilizing developing vascular networks, and transforming growth factor ß1 (TGF-ß1) has been shown to promote SMC differentiation from stem cells. Previously, our lab has developed a chemically modified fibrin-based hydrogel that induces endothelial cell (EC) phenotype and network formation from human mesenchymal stem cells (hMSCs) without exogenous cytokines. Additionally, we have shown that this hydrogel system is capable of releasing growth factors in a controlled manner. In the present work, the effects of TGF-ß1 on hMSCs in both monolayer and fibrin-based gel culture systems were demonstrated. The objective was to enhance SMC properties through TGF-ß1 signaling for vessel stability while maintaining EC gene expression and morphology. Proliferation was decreased with higher TGF-ß1 concentration in both monolayer and 3D gel cultures. EC genes were predominantly downregulated in the presence of TGF-ß1 in monolayer cultures, while SMC genes were generally upregulated. In fibrin-based gels, several SMC genes were significantly upregulated at high concentrations of TGF-ß1. Even at elevated TGF-ß1 concentrations, no significant differences were seen in EC genes for hMSCs in gels compared to controls. Network formation and growth occurred in PEGylated fibrin gels loaded with TGF-ß1 and were not significantly different from gels without loaded growth factor. Additionally, production of smooth muscle α-actin (SMA) was significantly increased in gels loaded with TGF-ß1. These results demonstrate a simultaneous response of hMSCs to both the 3D biomatrix and cytokine signaling cues.
RESUMO
Bone regeneration can be accelerated by utilizing mechanical stress and growth factors (GFs). However, a limited understanding exists regarding the response of preosteoblasts to tensile stress alone or with GFs. We measured cell proliferation and expression of heat-shock proteins (HSPs) and other bone-related proteins by preosteoblasts following cyclic tensile stress (1%-10% magnitude) alone or in combination with bone morphogenetic protein-2 (BMP-2) and transforming growth factor-ß1 (TGF-ß1). Tensile stress (3%) with GFs induced greater gene upregulation of osteoprotegerin (3.3 relative fold induction [RFI] compared to sham-treated samples), prostaglandin E synthase 2 (2.1 RFI), and vascular endothelial growth factor (VEGF) (11.5 RFI), compared with samples treated with stimuli alone or sham-treated samples. The most significant increases in messenger RNA expression occurred with GF addition to either static-cultured or tensile-loaded (1% elongation) cells for the following genes: HSP47 (RFI=2.53), cyclooxygenase-2 (RFI=72.52), bone sialoprotein (RFI=11.56), and TGF-ß1 (RFI=8.05). Following 5% strain with GFs, VEGF secretion increased 64% (days 3-6) compared with GF alone and cell proliferation increased 23% compared with the sham-treated group. GF addition increased osteocalcin secretion but decreased matrix metalloproteinase-9 significantly (days 3-6). Tensile stress and GFs in combination may enhance bone regeneration by initiating angiogenic and anti-osteoclastic effects and promote cell growth.
Assuntos
Proteína Morfogenética Óssea 2/farmacologia , Osso e Ossos/efeitos dos fármacos , Osso e Ossos/fisiologia , Osteoblastos/citologia , Estresse Mecânico , Engenharia Tecidual/métodos , Fator de Crescimento Transformador beta1/farmacologia , Animais , Matriz Óssea/efeitos dos fármacos , Matriz Óssea/metabolismo , Osso e Ossos/metabolismo , Linhagem Celular , Proliferação de Células/efeitos dos fármacos , Forma Celular/efeitos dos fármacos , Forma Celular/genética , Regulação da Expressão Gênica/efeitos dos fármacos , Proteínas de Choque Térmico/genética , Proteínas de Choque Térmico/metabolismo , Camundongos , Osteoblastos/efeitos dos fármacos , Osteoblastos/metabolismo , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Fatores de Tempo , Fator A de Crescimento do Endotélio Vascular/genética , Fator A de Crescimento do Endotélio Vascular/metabolismoRESUMO
Conditioning protocols involving mechanical stress independently or with chemical cues such as growth factors (GFs) possess significant potential to enhance bone regeneration. However, utilization of thermal stress conditioning alone or with GFs for bone therapy has been under-investigated. In this study, a preosteoblast cell line (MC3T3-E1) was exposed to treatment with water bath heating (44°C, 4 and 8 min) and osteoinductive GFs (bone morphogenetic protein-2 and transforming growth factor-ß1) individually or in combination to investigate whether these stimuli could promote induction of bone-related markers, an angiogenic factor, and heat shock proteins (HSPs). Cells remained viable when heating durations were less than 20 min at 40ºC, 16 min at 42ºC, and 10 min at 44ºC. Increasing heating duration at 44°C, promoted gene expression of HSPs, osteocalcin (OCN), and osteopontin (OPN) at 8 h post-heating (PH). Heating in combination with GFs caused the greatest gene induction of osteoprotegerin (OPG; 6.9- and 1.6-fold induction compared to sham-treated and GF only treated groups, respectively) and vascular endothelial growth factor (VEGF; 16.0- and 1.6-fold compared to sham and GF-only treated groups, respectively) at 8 h PH. Both heating and GFs independently suppressed the matrix metalloproteinase-9 (MMP-9) gene. GF treatment caused a more significant decrease in MMP-9 protein secretion to non-detectable levels compared to heating alone at 72 h PH. Secretion of OCN, OPN, and OPG increased with the addition of GFs but diminished with heating as measured by ELISA at 72 h PH. These results suggest that conditioning protocols utilizing heating and GFs individually or in combination can induce HSPs, bone-related proteins, and VEGF while also causing downregulation of osteoclastic activity, potentially providing a promising bone therapeutic strategy.
Assuntos
Proteína Morfogenética Óssea 2/farmacologia , Temperatura Alta , Osteoblastos/efeitos dos fármacos , Fator de Crescimento Transformador beta1/farmacologia , Animais , Linhagem Celular , Sobrevivência Celular/efeitos dos fármacos , Sobrevivência Celular/fisiologia , Regulação da Expressão Gênica/efeitos dos fármacos , Humanos , Camundongos , Osteoblastos/citologia , Osteoblastos/metabolismo , Osteocalcina/metabolismo , Osteopontina/metabolismo , Osteoprotegerina/genética , Osteoprotegerina/metabolismo , Fator A de Crescimento do Endotélio Vascular/genéticaRESUMO
Nanomedicine has great potential in biomedical applications, and specifically in regenerative medicine and vascular tissue engineering. Designing nanometer-sized therapeutic and diagnostic devices for tissue engineering applications is critical because cells experience and respond to stimuli on this spatial scale. For example, nanoscaffolds, including nanoscalestructured or nanoscale surface-modified vascular scaffolds, can influence cell alignment, adhesion, and differentiation to promote better endothelization. Furthermore, nanoscale contrast agents can be extended to the field of biomedical imaging to monitor and track stem cells to better understand the process of neovascularization. In addition, nanoscale systems capable of delivering biomolecules (e.g. peptides and angiogenic genes/proteins) can influence cell behavior, function, and phenotype to promote blood vessel regeneration. This review will focus on nanomedicine and nanoscale strategies applied to vascular tissue engineering. In particular, some of the latest research and potential applications pertaining to nanoscaffolds, biomedical imaging and cell tracking using nanoscale contrast agents, and nanodelivery systems of bioactive molecules applied to blood vessel regeneration will be discussed. In addition, the overlap between these three areas and their synergistic effects will be examined as related to vascular tissue engineering.
RESUMO
Tissue-engineered vascular grafts have been investigated as a substitute for prosthetic vascular grafts. The current scaffolds have several limitations due to weak mechanical properties in withstanding the pressure of blood vessel. A gel-spinning molding device including three-separate drivers that make a cylindrical shaft turn on its axis, orbit, and concurrently move up and down was developed for preparing seamless fibrous tubular scaffolds for vascular grafts. A seamless double-layered tubular scaffold, which was composed of an outer fibrous network and inner porous layer, was fabricated by using the device for the spinning of poly(L-lactide-co-caprolactone) (PLCL, 50:50) solution as a gel state on a rotating cylindrical shaft that had been dip-coated with the mixture of PLCL solution and NaCl particles. A scaffold that had an inner layer fabricated with 30% salts, below 20 mum in salt size, and more than 100 microm in thickness, was found to be optimal from a blood leakage test. The burst pressures of the scaffolds were more than 900 mmHg. The scaffolds exhibited 550-670% elongation-at-break. The measured circumferential and longitudinal tensile strengths of the scaffolds were 3.62 +/- 0.68 and 2.64 +/- 0.41 MPa, respectively. The suture retention strength of the scaffold was measured to be 7.68 +/- 0.75 N. These mechanically strong and elastic properties of the newly developed scaffolds provide an important basis for blood vessel tissue engineering.
Assuntos
Géis/química , Poliésteres/química , Engenharia Tecidual/instrumentação , Alicerces Teciduais/química , Elasticidade , Desenho de Equipamento , Teste de Materiais , Porosidade , ReologiaRESUMO
Biodegradable macroporous scaffolds have been developed for tissue-engineering applications. We fabricated and characterized a new tubular, macroporous, fibrous scaffold using a very elastic biodegradable co-polymer, poly(L-lactide-co-caprolactone) (PLCL, 5:5) in a gel-spinning process. A viscous PLCL solution was spun as a gel-phase under swirl-flow conditions and was subsequently fabricated to produce a tubular fibrous scaffold on a rotating cylindrical shaft in a methanol solution. The porosity and median pore size of the fibrous PLCL scaffolds were 55-75% and 120-150 microm, respectively, using a 5-10% PLCL solution. The use of a 7.5% (w/v) solution resulted in scaffolds with tensile strength and elastic modulus of 3.39 MPa and 1.22 MPa, respectively. The scaffolds exhibited 500-600% elongation-at-break. The tensile strength and modulus of fibrous PLCL scaffolds were proven to decrease on lowering the concentration of the PLCL spinning solution; however, the tensile strength and modulus of fibrous PLCL scaffolds, produced from 5% solutions, are approximately 4- and 5-times higher than those of extruded PLCL scaffolds. These properties indicated that the fibrous PLCL scaffolds were very elastic and mechanically strong. The scaffolds appeared to be well inter-connected between the pores as determined by SEM imaging analysis. In addition, the cell-seeding efficiency was 2-fold higher using gel-spun scaffolds than using extruded scaffolds. These results suggest that the gel-spun fibrous PLCL scaffold is an excellent matrix for vascular tissue-engineering applications.