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1.
Adv Exp Med Biol ; 1309: 257-276, 2021.
Artículo en Inglés | MEDLINE | ID: mdl-33782876

RESUMEN

The use of carbon-based nanomaterials (CNs) with outstanding properties has been rising in many scientific and industrial application fields. These CNs represent a tunable alternative for applications with biomolecules, which allow interactions in either covalent or noncovalent way. Diverse carbon-derived nanomaterial family exhibits unique features and has been widely exploited in various biomedical applications, including biosensing, diagnosis, cancer therapy, drug delivery, and tissue engineering. In this chapter, we aim to present an overview of CNs with a particular interest in intrinsic structural, electronic, and chemical properties. In particular, the detailed properties and features of CNs and its derivatives, including carbon nanotube (CNT), graphene, graphene oxide (GO), and reduced GO (rGO) are summarized. The interesting biomedical applications are also reviewed in order to offer an overview of the possible fields for scientific and industrial applications of CNs.


Asunto(s)
Nanoestructuras , Nanotubos de Carbono , Sistemas de Liberación de Medicamentos , Ingeniería de Tejidos
2.
J Biomed Nanotechnol ; 17(2): 242-252, 2021 Feb 28.
Artículo en Inglés | MEDLINE | ID: mdl-33785095

RESUMEN

Sericin, a silk protein, has a high potential for use as an extracellular matrix in tissue engineering applications. In this study, novel gelatin (GEL) and silk sericin (SS) were incorporated with a polyvinyl alcohol) PVA hydrogel nanocomposite (GEL-SS-PVA) scaffold that can be applied to repair cartilage. Glutaraldehyde was used as a cross-linking agent, with hydrochloric acid acting as an initiator. The microstructure characteristics of the obtained GEL-SS and GEL-SS-PVA scaffolds were also examined using FTIR and XRD spectra and their enhanced thermal stability was assessed by TGA. The blended GEL-SS and GEL-SS-PVA scaffolds were confirmed by SEM analysis to be highly porous with optimum pore sizes of 172 and 58 µm, respectively. Smaller pore sizes and improved uniformity were observed as the concentration of PVA in the GEL-SS-PVA scaffold increased. PVA decreased the tensile strength and elongation of the membranes but increased the modulus. Swelling studies showed high swellability and complete degradation in the presence of phosphate-buffered saline. Cytocompatibility of the GEL-SS-PVA scaffolds showed that these had the highest potential to promote cell proliferation as evaluated with standard microscopy using L929 fibroblasts. The prepared GEL-SS composite scaffold incorporated with the PVA hydrogel was implanted in full-thickness articular cartilage defects in rats. The repair effect of cartilage defects was observed and evaluated among the GEL-SS-PVA, GEL-SS, and control operation groups. The defects were almost completely repaired after 14 weeks in the GEL-SS-PVA group, thereby indicating that the GEL-SS-PVA composite had a favorable effect on articular cartilage defects in rat knee joint repair.


Asunto(s)
Cartílago Articular , Nanocompuestos , Sericinas , Animales , Gelatina , Hidrogeles , Articulación de la Rodilla , Alcohol Polivinílico , Ratas , Seda , Ingeniería de Tejidos , Andamios del Tejido
3.
Zhongguo Xiu Fu Chong Jian Wai Ke Za Zhi ; 35(3): 286-294, 2021 Mar 15.
Artículo en Chino | MEDLINE | ID: mdl-33719235

RESUMEN

Objective: To observe and compare the effects of peptides on the repair of rabbit skull defects through two different binding modes of non-covalent and covalent, and the combination of carboxyl (-COOH) and amino (-NH 2) groups with materials. Methods: Twenty-one 3-month-old male ordinary New Zealand white rabbits were numbered 1 to 42 on the left and right parietal bones. They were divided into 5 groups using a random number table, the control group (group A, 6 sides) and the material group 1, 2, 3, 4 (respectively group B, C, D, E, 9 sides in each group). All animals were prepared with 12-mm-diameter skull defect models, and bone morphogenetic protein 2 (BMP-2) non-covalently bound multiwalled carbon nanotubes (MWCNT)-COOH+poly ( L-lactide) (PLLA), BMP-2 non-covalently bound MWCNT-NH 2+PLLA, BMP-2 covalently bound MWCNT-COOH+PLLA, and BMP-2 covalently bound MWCNT-NH 2+PLLA were implanted into the defects of groups B, C, D, and E, respectively. At 4, 8, and 12 weeks after operation, the samples were taken for CT scanning and three-dimensional reconstruction, the ratio of bone tissue regeneration volume to total volume and bone mineral density were measured, and the histological observation of HE staining and Masson trichrome staining were performed to quantitatively analyze the volume ratio of new bone tissue. Results: CT scanning and three-dimensional reconstruction showed that with the extension of time, the defects in groups A-E were filled gradually, and the defect in group E was completely filled at 12 weeks after operation. HE staining and Masson trichrome staining showed that the volume of new bone tissue in each group gradually increased with time, and regenerated mature bone tissue appeared in groups D and E at 12 weeks after operation. Quantitative analysis showed that at 4, 8, and 12 weeks after operation, the ratio of bone tissue regeneration volume to total volume, bone mineral density, and the volume ratio of new bone tissue increased gradually over time; and at each time point, the above indexes increased gradually from group A to group E, and the differences between groups were significant ( P<0.05). Conclusion: Through covalent binding and using -NH 2 to bound peptides with materials, the best bone repair effect can be achieved.


Asunto(s)
Nanotubos de Carbono , Nanotubos de Péptidos , Animales , Proteína Morfogenética Ósea 2 , Regeneración Ósea , Masculino , Conejos , Cráneo , Ingeniería de Tejidos , Andamios del Tejido
4.
J Vis Exp ; (168)2021 02 12.
Artículo en Inglés | MEDLINE | ID: mdl-33645584

RESUMEN

Human skin equivalents (HSEs) are tissue engineered constructs that model epidermal and dermal components of human skin. These models have been used to study skin development, wound healing, and grafting techniques. Many HSEs continue to lack vasculature and are additionally analyzed through post-culture histological sectioning which limits volumetric assessment of the structure. Presented here is a straightforward protocol utilizing accessible materials to generate vascularized human skin equivalents (VHSE); further described are volumetric imaging and quantification techniques of these constructs. Briefly, VHSEs are constructed in 12 well culture inserts in which dermal and epidermal cells are seeded into rat tail collagen type I gel. The dermal compartment is made up of fibroblast and endothelial cells dispersed throughout collagen gel. The epidermal compartment is made up of keratinocytes (skin epithelial cells) that differentiate at the air-liquid interface. Importantly, these methods are customizable based on needs of the researcher, with results demonstrating VHSE generation with two different fibroblast cell types: human dermal fibroblasts (hDF) and human lung fibroblasts (IMR90s). VHSEs were developed, imaged through confocal microscopy, and volumetrically analyzed using computational software at 4- and 8-week timepoints. An optimized process to fix, stain, image, and clear VHSEs for volumetric examination is described. This comprehensive model, imaging, and analysis techniques are readily customizable to the specific research needs of individual labs with or without prior HSE experience.


Asunto(s)
Neovascularización Fisiológica , Piel Artificial , Piel/irrigación sanguínea , Ingeniería de Tejidos/métodos , Animales , Biomarcadores/metabolismo , Células Cultivadas , Colágeno/metabolismo , Dermis/metabolismo , Epidermis/metabolismo , Técnica del Anticuerpo Fluorescente , Humanos , Imagenología Tridimensional , Imagen Óptica , Permeabilidad , Ratas , Coloración y Etiquetado , Suspensiones
5.
Methods Mol Biol ; 2269: 175-201, 2021.
Artículo en Inglés | MEDLINE | ID: mdl-33687680

RESUMEN

Bench-to-bedside axis of therapeutic product development is currently being oriented towards minimum invasiveness on both ends-not only clinical application but harvesting of the starting biological material as well. This is particularly relevant for Advanced Therapy Medicinal Products and their specific legislative requirements, even more so in skin regeneration. It is precisely the skin equivalents and grafts that benefit from the minimum-to-noninvasive approach to a noteworthy extent, taking in account the sensitive nature of both skin harvesting and grafting.This chapter includes protocols for two separate steps of generating skin equivalent from the cells cultured from hair follicle outer root sheath. The first step is a non-pigmented epidermal equivalent generated from human keratinocytes from the outer root sheath named non-pigmented epidermal graft. The second step consists of co-cultivating human keratinocytes and human melanocytes from the outer root sheath, hereby producing a pigmented epidermal graft.


Asunto(s)
Dermis/metabolismo , Fibroblastos/metabolismo , Folículo Piloso/metabolismo , Queratinocitos/metabolismo , Melanocitos/metabolismo , Ingeniería de Tejidos , Técnicas de Cocultivo , Dermis/citología , Fibroblastos/citología , Folículo Piloso/citología , Humanos , Queratinocitos/citología , Melanocitos/citología
6.
Methods Mol Biol ; 2269: 245-254, 2021.
Artículo en Inglés | MEDLINE | ID: mdl-33687684

RESUMEN

Peripheral nerves have a limited ability to regenerate and current clinical approaches involving microsurgery give suboptimal recovery. Engineered tissues using aligned cellular collagen hydrogels can be used as in vitro models through the incorporation of human Schwann cells. However, primary human Schwann cells are difficult to obtain and can be challenging to culture. The ability to generate Schwann cells from human-induced pluripotent stem cells (hiPSCs) provides a more reliable cell source for modeling peripheral nerve tissue. Here, we describe protocols for generating hiPSC-derived Schwann cells and incorporating them into 3D engineered tissue culture models for peripheral nerve research.


Asunto(s)
Diferenciación Celular , Células Madre Pluripotentes Inducidas/metabolismo , Regeneración Nerviosa , Nervios Periféricos/metabolismo , Células de Schwann/metabolismo , Ingeniería de Tejidos , Humanos , Células Madre Pluripotentes Inducidas/citología , Nervios Periféricos/citología
7.
J Vis Exp ; (168)2021 02 18.
Artículo en Inglés | MEDLINE | ID: mdl-33682863

RESUMEN

Three-dimensional (3D) in vitro models of skeletal muscle are a valuable advancement in biomedical research as they afford the opportunity to study skeletal muscle reformation and function in a scalable format that is amenable to experimental manipulations. 3D muscle culture systems are desirable as they enable scientists to study skeletal muscle ex vivo in the context of human cells. 3D in vitro models closely mimic aspects of the native tissue structure of adult skeletal muscle. However, their universal application is limited by the availability of platforms that are simple to fabricate, cost and user-friendly, and yield relatively high quantities of human skeletal muscle tissues. Additionally, since skeletal muscle plays an important functional role that is impaired over time in many disease states, an experimental platform for microtissue studies is most practical when minimally invasive calcium transient and contractile force measurements can be conducted directly within the platform itself. In this protocol, the fabrication of a 96-well platform known as 'MyoTACTIC', and en masse production of 3D human skeletal muscle microtissues (hMMTs) is described. In addition, the methods for a minimally invasive application of electrical stimulation that enables repeated measurements of skeletal muscle force and calcium handling of each microtissue over time are reported.


Asunto(s)
Salud , Músculo Esquelético/fisiología , Ingeniería de Tejidos , Calcio/metabolismo , Diferenciación Celular , Línea Celular Transformada , Dimetilpolisiloxanos/química , Estimulación Eléctrica , Humanos , Contracción Muscular/fisiología , Fibras Musculares Esqueléticas/citología , Mioblastos/citología
8.
Carbohydr Polym ; 260: 117765, 2021 May 15.
Artículo en Inglés | MEDLINE | ID: mdl-33712123

RESUMEN

Chitosan (CS) combined with hydroxyapatite (HA) was injected into a composite braid, and a hierarchical pore structure scaffold was obtained by freeze drying and cold atmospheric plasma (CAP) technology. The CS/HA/braid scaffold with hierarchical pore structure was analyzed and characterized by scanning electronic microscopy, Fourier transform infrared spectroscopy, true color confocal microscopy, improved liquid replacement method, and phosphate buffer solution immersion. The mechanical properties and degradation ability of the scaffold were evaluated through compression test and degradation test. Results showed that HA addition endowed the core of the scaffold with macroscopic pore sizes of 80-180 µm, and CAP treatment endowed the shell of the scaffold with microscopic pore sizes ≤10 µm. All scaffolds exhibited high porosity and swelling rates of ≥80 % and ≥300 %, respectively. The scaffold with a hierarchical pore structure had good mechanical properties and twice the degradation rate. In addition, the treated scaffold precipitated intact spherical HA crystals. Under the synergistic effect of HA and CAP treatment, scaffolds achieved 277.6 % cell viability compared with pure CS scaffold. Overall, this method was feasible for preparing bone scaffolds with hierarchical pore structure for potential bone tissue engineering.


Asunto(s)
Quitosano/química , Durapatita/química , Ingeniería de Tejidos , Andamios del Tejido/química , Animales , Materiales Biocompatibles/química , Materiales Biocompatibles/farmacología , Línea Celular , Supervivencia Celular/efectos de los fármacos , Fuerza Compresiva , Liofilización , Ratones , Porosidad
9.
Carbohydr Polym ; 260: 117768, 2021 May 15.
Artículo en Inglés | MEDLINE | ID: mdl-33712126

RESUMEN

Tissue engineering and regenerative medicine have entered a new stage of development by the recent progress in biology, material sciences, and particularly an emerging additive manufacturing technique, three-dimensional (3D) printing. 3D printing is an advanced biofabrication technique which can generate patient-specific scaffolds with highly complex geometries while hosting cells and bioactive agents to accelerate tissue regeneration. Chitosan hydrogels themselves have been widely used for various biomedical applications due to its abundant availability, structural features and favorable biological properties; however, the 3D printing of chitosan-based hydrogels is still under early exploration. Therefore, 3D printing technologies represent a new avenue to explore the potential application of chitosan as an ink for 3D printing, or as a coating on other 3D printed scaffolds. The combination of chitosan-based hydrogels and 3D printing holds much promise in the development of next generation biomedical implants.


Asunto(s)
Quitosano/química , Hidrogeles/química , Impresión Tridimensional , Materiales Biocompatibles/química , Humanos , Medicina Regenerativa , Ingeniería de Tejidos
10.
Carbohydr Polym ; 260: 117780, 2021 May 15.
Artículo en Inglés | MEDLINE | ID: mdl-33712136

RESUMEN

In this study, we prepared a biomimetic hyaluronic acid oligosaccharides (oHAs)-based composite scaffold to develop a bone tissue-engineered scaffold for stimulating osteogenesis and endothelialization. The functional oHAs products were firstly synthesized, namely collagen/hyaluronic acid oligosaccharides/hydroxyapatite (Col/oHAs/HAP), chitosan/hyaluronic acid oligosaccharides (CTS/oHAs), and then uniformly distributed in poly (lactic-co-glycolic acid) (PLGA) solution followed by freeze-drying to obtain three-dimensional interconnected scaffolds as temporary templates for bone regeneration. The morphology, physicochemical properties, compressive strength, and degradation behavior of the fabricated scaffolds, as well as in vitro cell responses seeded on these scaffolds and in vivo biocompatibility, were investigated to evaluate the potential for bone tissue engineering. The results indicated that the oHAs-based scaffolds can promote the attachment of endothelial cells, facilitate the osteogenic differentiation of MC3T3-E1 and BMSCs, and have ideal biocompatibility and tissue regenerative capacity, suggesting their potential to serve as alternative candidates for bone tissue engineering applications.


Asunto(s)
Materiales Biocompatibles/química , Quitosano/química , Colágeno/química , Ingeniería de Tejidos , Animales , Materiales Biocompatibles/farmacología , Diferenciación Celular/efectos de los fármacos , Línea Celular , Proliferación Celular/efectos de los fármacos , Durapatita/química , Ácido Hialurónico/química , Células Madre Mesenquimatosas/citología , Células Madre Mesenquimatosas/metabolismo , Ratones , Oligosacáridos/química , Osteoblastos/citología , Osteoblastos/metabolismo , Osteogénesis/efectos de los fármacos , Copolímero de Ácido Poliláctico-Ácido Poliglicólico/química , Andamios del Tejido/química
11.
Molecules ; 26(5)2021 Feb 24.
Artículo en Inglés | MEDLINE | ID: mdl-33668087

RESUMEN

Stereolithography is a useful additive manufacturing technique for the production of scaffolds for tissue engineering. Here we present a tuneable, easy-to-manufacture, photocurable resin for use in stereolithography, based on the widely used biomaterial, poly(caprolactone) (PCL). PCL triol was methacrylated to varying degrees and mixed with photoinitiator to produce a photocurable prepolymer resin, which cured under UV light to produce a cytocompatible material. This study demonstrates that poly(caprolactone) methacrylate (PCLMA) can be produced with a range of mechanical properties and degradation rates. By increasing the degree of methacrylation (DM) of the prepolymer, the Young's modulus of the crosslinked PCLMA could be varied from 0.12-3.51 MPa. The accelerated degradation rate was also reduced from complete degradation in 17 days to non-significant degradation in 21 days. The additive manufacturing capabilities of the resin were demonstrated by the production of a variety of different 3D structures using micro-stereolithography. Here, ß-carotene was used as a novel, cytocompatible photoabsorber and enabled the production of complex geometries by giving control over cure depth. The PCLMA presented here offers an attractive, tuneable biomaterial for the production of tissue engineering scaffolds for a wide range of applications.


Asunto(s)
Materiales Biocompatibles/química , Poliésteres/química , Resinas Sintéticas/química , Estereolitografía , Ingeniería de Tejidos , Andamios del Tejido/química , Materiales Biocompatibles/síntesis química , Estructura Molecular , Procesos Fotoquímicos , Poliésteres/síntesis química , Resinas Sintéticas/síntesis química , beta Caroteno/química
12.
Ann R Coll Surg Engl ; 103(4): 245-249, 2021 Apr.
Artículo en Inglés | MEDLINE | ID: mdl-33682428

RESUMEN

Soft tissue reconstruction remains a continuing challenge for plastic and reconstructive surgeons. Standard methods of reconstruction such as local tissue transfer and free autologous tissue transfer are successful in addressing soft tissue cover, yet they do not come without the additional morbidity of donor sites. Autologous fat transfer has been used in reconstruction of soft tissue defects in different branches of plastic surgery, specifically breast and facial defect reconstruction, while further maintaining a role in body contouring procedures. Current autologous fat transfer techniques come with the drawbacks of donor-site morbidity and, more significantly, resorption of large amounts of fat. Advancement in tissue engineering has led to the use of engineered adipose tissue structures based on adipose-derived stem cells. This enables a mechanically similar reconstruct that is abundantly available. Cosmetic and mechanical similarity with native tissue is the main clinical goal for engineered adipose tissue. Development of novel techniques in the availability of natural tissue is an exciting prospect; however, it is important to investigate the potential of cell sources and culture strategies for clinical applications. We review these techniques and their applications in plastic surgery.


Asunto(s)
Tejido Adiposo/trasplante , Trasplante de Células Madre Mesenquimatosas , Células Madre Mesenquimatosas , Procedimientos Quirúrgicos Reconstructivos/métodos , Ingeniería de Tejidos/métodos , Tejido Adiposo/citología , Humanos
13.
Nat Methods ; 18(2): 119, 2021 02.
Artículo en Inglés | MEDLINE | ID: mdl-33542506
14.
Methods Mol Biol ; 2235: 127-137, 2021.
Artículo en Inglés | MEDLINE | ID: mdl-33576974

RESUMEN

Human pericytes are a perivascular cell population with mesenchymal stem cell properties, present in all vascularized tissues. Human pericytes have a distinct immunoprofile, which may be leveraged for purposes of cell purification. Adipose tissue is the most commonly used cell source for human pericyte derivation. Pericytes can be isolated by FACS (fluorescence-activated cell sorting), most commonly procured from liposuction aspirates. Pericytes have clonal multilineage differentiation potential, and their potential utility for bone regeneration has been described across multiple animal models. The following review will discuss in vivo methods for assessing the bone-forming potential of purified pericytes. Potential models include (1) mouse intramuscular implantation, (2) mouse calvarial defect implantation, and (3) rat spinal fusion models. In addition, the presented surgical protocols may be used for the in vivo analysis of other osteoprogenitor cell types.


Asunto(s)
Células de la Médula Ósea/metabolismo , Pericitos/metabolismo , Ingeniería de Tejidos/métodos , Tejido Adiposo/citología , Animales , Células de la Médula Ósea/citología , Regeneración Ósea/fisiología , Diferenciación Celular/efectos de los fármacos , Diferenciación Celular/fisiología , Línea Celular , Separación Celular/métodos , Humanos , Células Madre Mesenquimatosas/citología , Ratones , Osteogénesis/fisiología , Pericitos/citología , Ratas
15.
Methods Mol Biol ; 2273: 111-129, 2021.
Artículo en Inglés | MEDLINE | ID: mdl-33604848

RESUMEN

Tissue engineering provides unique opportunities for disease modeling, drug testing, and regenerative medicine applications. The use of cell-seeded scaffolds to promote tissue development is the hallmark of the tissue engineering. Among the different types of scaffolds (derived from either natural or synthetic polymers) used in the field, the use of decellularized tissues/organs is specifically attractive. The decellularization process involves the removal of native cells from the original tissue, allowing for the preservation of the three-dimensional (3D) macroscopic and microscopic structures of the tissue and extracellular matrix (ECM) composition. Following recellularization, the resulting scaffold provides the seeded cells with the appropriate biological signals and mechanical properties of the original tissue. Here, we describe different methods to create viable scaffolds from decellularized heart and liver as useful tools to study and exploit ECM biological key factors for the generation of engineered tissues with enhanced regenerative properties.


Asunto(s)
Dermis Acelular/metabolismo , Medicina Regenerativa/métodos , Ingeniería de Tejidos/métodos , Animales , Matriz Extracelular/química , Corazón/crecimiento & desarrollo , Hepatocitos/citología , Hígado/crecimiento & desarrollo , Miocitos Cardíacos/citología , Conejos
16.
Methods Mol Biol ; 2273: 139-149, 2021.
Artículo en Inglés | MEDLINE | ID: mdl-33604850

RESUMEN

Ovarian failure is the most common cause of infertility and affects about 1% of young women. One innovative strategy to restore ovarian function may be represented by the development of a bioprosthetic ovary, obtained through the combination of tissue engineering and regenerative medicine.We here describe the two main steps required for bioengineering the ovary and for its ex vivo functional reassembling. The first step aims at producing a 3D bioscaffold, which mimics the natural ovarian milieu in vitro. This is obtained with a whole organ decellularization technique that allows the maintenance of microarchitecture and biological signals of the original tissue. The second step involves the use of magnetic activated cell sorting (MACS) to isolate purified female germline stem cells (FGSCs). These cells are able to differentiate in ovarian adult mature cells, when subjected to specific stimuli, and can be used them to repopulate ovarian decellularized bioscaffolds. The combination of the two techniques represents a powerful tool for in vitro recreation of a bioengineered ovary that may constitute a promising solution for hormone and fertility function restoring. In addition, the procedures here described allow for the creation of a suitable 3D platform with useful applications both in toxicological and transplantation studies.


Asunto(s)
Células Madre Oogoniales/trasplante , Ovario/crecimiento & desarrollo , Ingeniería de Tejidos/métodos , Animales , Bioingeniería/métodos , Ingeniería Biomédica , Técnicas de Cultivo de Célula/métodos , Matriz Extracelular/metabolismo , Femenino , Fertilidad , Humanos , Células Madre Oogoniales/metabolismo , Organoides/crecimiento & desarrollo , Medicina Regenerativa , Porcinos , Andamios del Tejido/química
17.
Int J Mol Sci ; 22(3)2021 Jan 26.
Artículo en Inglés | MEDLINE | ID: mdl-33530458

RESUMEN

A high-throughput drug screen identifies potentially promising therapeutics for clinical trials. However, limitations that persist in current disease modeling with limited physiological relevancy of human patients skew drug responses, hamper translation of clinical efficacy, and contribute to high clinical attritions. The emergence of induced pluripotent stem cell (iPSC) technology revolutionizes the paradigm of drug discovery. In particular, iPSC-based three-dimensional (3D) tissue engineering that appears as a promising vehicle of in vitro disease modeling provides more sophisticated tissue architectures and micro-environmental cues than a traditional two-dimensional (2D) culture. Here we discuss 3D based organoids/spheroids that construct the advanced modeling with evolved structural complexity, which propels drug discovery by exhibiting more human specific and diverse pathologies that are not perceived in 2D or animal models. We will then focus on various central nerve system (CNS) disease modeling using human iPSCs, leading to uncovering disease pathogenesis that guides the development of therapeutic strategies. Finally, we will address new opportunities of iPSC-assisted drug discovery with multi-disciplinary approaches from bioengineering to Omics technology. Despite technological challenges, iPSC-derived cytoarchitectures through interactions of diverse cell types mimic patients' CNS and serve as a platform for therapeutic development and personalized precision medicine.


Asunto(s)
Enfermedades del Sistema Nervioso Central/tratamiento farmacológico , Descubrimiento de Drogas/métodos , Células Madre Pluripotentes Inducidas/citología , Células Madre Pluripotentes Inducidas/efectos de los fármacos , Ingeniería de Tejidos/métodos , Animales , /patología , Enfermedades del Sistema Nervioso Central/patología , Descubrimiento de Drogas/instrumentación , Evaluación Preclínica de Medicamentos/instrumentación , Evaluación Preclínica de Medicamentos/métodos , Humanos , Células Madre Pluripotentes Inducidas/patología , Dispositivos Laboratorio en un Chip , Organoides/citología , Organoides/efectos de los fármacos , Organoides/patología , Ingeniería de Tejidos/instrumentación , Infección por el Virus Zika/tratamiento farmacológico , Infección por el Virus Zika/patología
18.
Int J Nanomedicine ; 16: 989-1000, 2021.
Artículo en Inglés | MEDLINE | ID: mdl-33633447

RESUMEN

Background: Under certain conditions, the physiological repair of connective tissues might fail to restore the original structure and function. Optimized engineered connective tissues (ECTs) with biophysical properties adapted to the target tissue could be used as a substitution therapy. This study aimed to investigate the effect of ECT enforcement by a complex of multiwall carbon nanotubes with chitosan (C-MWCNT) to meet in vivo demands. Materials and Methods: ECTs were constructed from human foreskin fibroblasts (HFF-1) in collagen type I and enriched with the three different percentages 0.025, 0.05 and 0.1% of C-MWCNT. Characterization of the physical properties was performed by biomechanical studies using unidirectional strain. Results: Supplementation with 0.025% C-MWCNT moderately increased the tissue stiffness, reflected by Young's modulus, compared to tissues without C-MWCNT. Supplementation of ECTs with 0.1% C-MWCNT reduced tissue contraction and increased the elasticity and the extensibility, reflected by the yield point and ultimate strain, respectively. Consequently, the ECTs with 0.1% C-MWCNT showed a higher resilience and toughness as control tissues. Fluorescence tissue imaging demonstrated the longitudinal alignment of all cells independent of the condition. Conclusion: Supplementation with C-MWCNT can enhance the biophysical properties of ECTs, which could be advantageous for applications in connective tissue repair.


Asunto(s)
Quitosano/farmacología , Tejido Conectivo/fisiología , Nanotubos de Carbono/química , Ingeniería de Tejidos , Animales , Fenómenos Biomecánicos , Bovinos , Línea Celular , Quitosano/química , Módulo de Elasticidad , Fibroblastos/efectos de los fármacos , Humanos
19.
J Vis Exp ; (167)2021 01 14.
Artículo en Inglés | MEDLINE | ID: mdl-33522507

RESUMEN

The cardiovascular system is a key player in human physiology, providing nourishment to most tissues in the body; vessels are present in different sizes, structures, phenotypes, and performance depending on each specific perfused tissue. The field of tissue engineering, which aims to repair or replace damaged or missing body tissues, relies on controlled angiogenesis to create a proper vascularization within the engineered tissues. Without a vascular system, thick engineered constructs cannot be sufficiently nourished, which may result in cell death, poor engraftment, and ultimately failure. Thus, understanding and controlling the behavior of engineered blood vessels is an outstanding challenge in the field. This work presents a high-throughput system that allows for the creation of organized and repeatable vessel networks for studying vessel behavior in a 3D scaffold environment. This two-step seeding protocol shows that vessels within the system react to the scaffold topography, presenting distinctive sprouting behaviors depending on the compartment geometry in which the vessels reside. The obtained results and understanding from this high throughput system can be applied in order to inform better 3D bioprinted scaffold construct designs, wherein fabrication of various 3D geometries cannot be rapidly assessed when using 3D printing as the basis for cellularized biological environments. Furthermore, the understanding from this high throughput system may be utilized for the improvement of rapid drug screening, the rapid development of co-cultures models, and the investigation of mechanical stimuli on blood vessel formation to deepen the knowledge of the vascular system.


Asunto(s)
Vasos Sanguíneos/crecimiento & desarrollo , Neovascularización Fisiológica , Ingeniería de Tejidos/métodos , Andamios del Tejido/química , Actinas/metabolismo , Biomarcadores/metabolismo , Movimiento Celular , Células Cultivadas , Técnicas de Cocultivo , Células Endoteliales/efectos de los fármacos , Fibronectinas/farmacología , Técnica del Anticuerpo Fluorescente , Humanos , Impresión Tridimensional , Imagen de Lapso de Tiempo
20.
Nat Commun ; 12(1): 1031, 2021 02 15.
Artículo en Inglés | MEDLINE | ID: mdl-33589620

RESUMEN

The application of physical stimuli to cell cultures has shown potential to modulate multiple cellular functions including migration, differentiation and survival. However, the relevance of these in vitro models to future potential extrapolation in vivo depends on whether stimuli can be applied "externally", without invasive procedures. Here, we report on the fabrication and exploitation of dynamic additive-manufactured Janus scaffolds that are activated on-command via external application of ultrasounds, resulting in a mechanical nanovibration that is transmitted to the surrounding cells. Janus scaffolds were spontaneously formed via phase-segregation of biodegradable polycaprolactone (PCL) and polylactide (PLA) blends during the manufacturing process and behave as ultrasound transducers (acoustic to mechanical) where the PLA and PCL phases represent the active and backing materials, respectively. Remote stimulation of Janus scaffolds led to enhanced cell proliferation, matrix deposition and osteogenic differentiation of seeded human bone marrow derived stromal cells (hBMSCs) via formation and activation of voltage-gated calcium ion channels.


Asunto(s)
Plásticos Biodegradables/farmacología , Mecanotransducción Celular , Células Madre Mesenquimatosas/efectos de los fármacos , Poliésteres/farmacología , Andamios del Tejido , Plásticos Biodegradables/química , Regeneración Ósea/genética , Huesos/citología , Huesos/metabolismo , Canales de Calcio Activados por la Liberación de Calcio/fisiología , Diferenciación Celular/efectos de los fármacos , Línea Celular , Proliferación Celular/efectos de los fármacos , Humanos , Células Madre Mesenquimatosas/citología , Células Madre Mesenquimatosas/metabolismo , Poliésteres/química , Impresión Tridimensional , Ingeniería de Tejidos/métodos , Ondas Ultrasónicas
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