Nanotechnology In Regenerative Medicine Methods And Protocols Pdf

nanotechnology in regenerative medicine methods and protocols pdf

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DNA nanotechnology is the design and manufacture of artificial nucleic acid structures for technological uses. In this field, nucleic acids are used as non-biological engineering materials for nanotechnology rather than as the carriers of genetic information in living cells.

Metrics details. In recent years, stem cell therapy has become a very promising and advanced scientific research topic. The development of treatment methods has evoked great expectations.

Exploring innovative solutions to improve the healthcare of the aging and diseased population continues to be a global challenge. Among a number of strategies toward this goal, tissue engineering and regenerative medicine TERM has gradually evolved into a promising approach to meet future needs of patients. TERM has recently received increasing attention in Asia, as evidenced by the markedly increased number of researchers, publications, clinical trials, and translational products.

Stem cells: past, present, and future

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In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript. The worldwide incidence of bone disorders and conditions has been increasing. Bone is a nanomaterials composed of organic mainly collagen and inorganic mainly nano-hydroxyapatite components, with a hierarchical structure ranging from nanoscale to macroscale.

In consideration of the serious limitation in traditional therapies, nanomaterials provide some new strategy in bone regeneration. Nanostructured scaffolds provide a closer structural support approximation to native bone architecture for the cells and regulate cell proliferation, differentiation, and migration, which results in the formation of functional tissues. In this article, we focused on reviewing the classification and design of nanostructured materials and nanocarrier materials for bone regeneration, their cell interaction properties, and their application in bone tissue engineering and regeneration.

Furthermore, some new challenges about the future research on the application of nanomaterials for bone regeneration are described in the conclusion and perspectives part. Nowadays more and more bone diseases such as bone infections, bone tumors, and bone loss need for bone regeneration. Bone tissue engineering is a complex and dynamic process that initiates with migration and recruitment of osteoprogenitor cells followed by their proliferation, differentiation, matrix formation along with remodeling of the bone.

Researches on bone tissue engineering over the past decades have inspired innovation in novel materials, processing techniques, performance evaluation, and applications. Significant progress has been made toward scaffold materials for structural support for desired osteogenesis and angiogenesis abilities. Bioresorbable scaffolds with controlled porosity and tailored properties are possible today due to innovation in scaffold fabrication using advanced technologies.

Natural bone derives its unique combination of mechanical properties from an architectural design that spans nanoscale to macroscopic dimensions, with precisely and carefully engineered interfaces. Many different groups have tried to manipulate the mechanical properties e. The ECM is composed of a wide spectrum of structural proteins and polysaccharides that span over different length scales, with strands of collagen fibrils dominating at the nanometer level, with a diameter between 35 and 60 nm and a length that can extend over the micron range.

The advent of sophisticated small-scale technologies has now made it possible for researchers to fabricate platforms that can be used to gain valuable insights into stem cell biomechanics. Nonetheless, despite the significance of stem cell mechanobiology, how mechanical stimuli regulates the behaviors of stem cells both in vivo and ex vivo have yet to be fully understood. To better mimic the nanostructure in natural ECM, over the past decade, scaffolds manufactured from nanofibers, nanotubes, nanoparticles, and hydrogel have recently emerged as promising candidates in producing scaffolds that resemble the ECM and efficiently replace defective tissues.

Following the approach of scaffolding as a way of temporarily mimicking the ECM of bone, it is necessary to look at the chemical, mechanical, and structural properties of bone. Bone is a sophisticated composite on different hierarchical levels, as shown in Figure 1. Bone tissue consists of two main parts, a compact shell called cortical bone and a porous core called spongiosa or trabecular bone Figure 1a.

Cortical bone is composed of repeating osteon units, whereas the cancellous bone is made of an interconnecting framework of trabeculae with bone marrow-filled free spaces. These trabeculae and osteon units are composed of collagen fibers and calcium phosphate crystals.

The collagen fibrils include a 67 nm periodicity and 40 nm gaps between collagen molecules Figure 1b. The microstructure and nanostructure of bone and the nanostructured material used in bone regeneration. The tertiary structure of collagen fibrils includes a 67 nm periodicity and 40 nm gaps between collagen molecules.

The hydroxyapatite HA crystals are embedded in these gaps between collagen molecules and increase the rigidity of the bone. Nanostructures with features of nanopattern d , nanofibers e , nanotubers f , nanopores g , nanospheres h , and nanocomposites i with structural components with a feature size in the nanoscale. Bone regeneration requires four components: a morphogenetic signal, responsive host cells that will respond to the signal, a suitable carrier of this signal that can deliver it to specific sites then serve as a scaffold for the growth of the responsive host cells, and a viable, well-vascularized host bed.

Materials used as bone tissue-engineered scaffolds may be injectable or rigid, with the latter requiring an operative implantation procedure. As a bone tissue engineering, the ideal bone tissue scaffolds should be osteoconductive, osteoinductive, and osteogenic. Osteoinductive scaffolds offer physical and biochemical factor to induce stem cells toward osteoblastic lineage. Osteogenic scaffolds contain osteogenic stem cells for bone regeneration.

In a word, bone tissue engineering utilizes a biomimetic strategy which includes suitable scaffolds, biochemical and physical stimuli, stem cells, vascularization, and recapitulating the hierarchical organization of natural ECM to create functional bone tissues. These biomimetic efforts include choosing biomaterials that are present in native bone e.

Conventional tissue engineering scaffolds have used various pore-forming methods to recreate the macroscale and microscale properties of native tissues, but the nanoscale structures and properties were neglected.

However, the nanoscale structures are crucial to regulating cell functions, such as proliferation, migration, differentiation, and the formation of ECM. To simulate the hierarchical organization of natural ECM, one important strategy is to build nanoscale and microscale features in the three-dimensional 3D scaffolds design. The commonly accepted definition of nanomaterials refers to materials with clearly defined features between 1 and nm, such as nanopattern, 15 nanofibers, 16 nanotubers, 17 nanopores, 18 nanospheres, 19 and nanocomposites 20 , 21 Figure 1d—i.

The construction of synthetic ECMs inspired by tissue-specific niches for programmed stem cell fate and response, such as proliferation and differentiation, is a topic of interest in the field of tissue regeneration. Using nanogrooved matrices mimicking the native tissues, Kim et al. The effect of nano-topographical density on the osteo- or neurogenesis of hMSCs was significant at the and nanogrooved patterns, but not significant at nanogrooved pattern compared to that at the flat substrate.

It is demonstrated the effects of nano-topographical density on the morphology and differentiation of mesenchymal stem cells. The formation of cytoskeleton is necessary for the shape effect on the stem cell differentiation and that the Rho-associated protein kinase ROCK -pathway-related cell tension is responsible for this effect in the case of osteogenesis even in growth medium.

The adipogenic differentiation does not seem to be simply negatively related to cell tension, and otherwise the adipogenic fraction might be minimum in the case of large aspect ratio of cells with the highest cell tension.

Schematic depictions of representative nanotopography geometries. Three basic nanotopography geometries include nanogrooves a , nanopost array b , and nanopit array c. The speculative pathways d for cell-shape-directed osteogenic and adipogenic differentiations of MSCs were examined in growth medium.

Jangho Kim supports the notion that multiscale hierarchical topography can be used as an efficient strategy for the design and manipulation of synthetic ECMs for stem cell-based bone regeneration. A type of synthetic ECM comprised of hierarchically multiscale structures could provide native ECM-like topographical cues for controlling the adhesion and differentiation of hMSCs. Interestingly, the platform that integrates hMSCs into the multiscale hierarchical PLGA patch showed the potential to regenerate the bone tissues without complex surgical treatments.

They work provides insight into the design and manipulation of functional engineered constructs using multi-scale hierarchical topography-based substrates for various biomedical applications, including stem cell therapy and tissue engineering.

Cell shape, in particular, the degree of cell spreading reflected in the cell area, is known to influence cell fate decisions of hMSC. Since cell area was shown to be regulated by the density of nanoposts, Ahn et al. The finding suggested that the nanoposts density might be capable of directly regulating cytoskeletal stiffness and the dynamic changes in stiffness correlated with the differentiation of hMSC into osteogenic or adipogenic lineages.

Using spatially ordered and disordered arrays of nanopatterned c-RGDfK peptide with well-defined interpattern distances that ranged from 55 to nm against a non-adhesive background, Huang et al. As is known, synthetically nanofabricated topography can also influence cell morphology, alignment, adhesion, migration, proliferation, and cytoskeleton organization.

Increased bone nodule formation was also evident in hMSCs cultured on these substrates relative to substrates with either completely ordered or completely random features. The results from the studies demonstrated the potential of nanotopography to direct cell fate.

Furthermore, the complementary findings of hMSCs cultured on nanogratings and ordered-disordered nanopits suggested the potential for selective, controllable differentiation based solely on the geometry of the nanotopographic substrate. Collectively, a few common observations can be drawn from the before mentioned studies of the mechano-sensitivity of stem cells. All the studies have explicitly or implicitly suggested the involvement of cytoskeleton contractility in regulating the mechanosensitivity of stem cells, suggesting the importance of the force balance along the mechanical axis of the ECM—integrin—cytoskeleton linkage and their regulation by the mechanical signals in the stem cell niche Figure 2d.

The principle of electrospinning is that an electric field is used to overcome the surface tension of a polymer solution to shoot a jet of liquid out of a needle toward a conducting collector. Many parameters affect this process including polymer properties, solvent properties, solution flow rate, voltage, distance from the needle to the collector, and polymer concentration, among others.

Generally, there are two types of polymers that are chosen: synthetic polymers or natural polymers. However, these polymers lack bioactivity and special care needs to be taken to ensure that newly synthesized polymers are biocompatible.

Many natural polymers, on the other hand, have inherent bioactivity with peptide sequences that affect cell adhesion, proliferation, and differentiation. Collagen, gelatin, silk, and chitosan, among others, are commonly used natural polymers for scaffold fabrication, but care must be taken to prevent denaturation when proteins are used.

Since both synthetic and natural polymers have advantages and disadvantages, research has progressed to fabricate hybrid scaffolds in an effort to maximize the benefits of both.

Yang et al. This novel hybrid scaffold takes advantage of the physical properties of the synthetic polymer and the bioactivity of the natural polymer while minimizing the disadvantages of both.

To develop biomimetic bone tissue engineering scaffolds for the repair of critical-sized calvarial defect, and growth factors can be incorporated into the polymer to create a controlled delivery system Figure 3. Li et al. The in vitro studies showed that the bioactivity of DEX and BMP-2 was preserved in the dual-drug-loaded nanofiber scaffold, and a sequential release pattern in which most of the DEX was released in the original 8 days and the BMP-2 release lasted up to 35 days was achieved.

The in vitro osteogenesis study demonstrated that the drug-loaded groups exhibited a strong ability to induce differentiation toward osteoblasts. In vivo osteogenesis studies also revealed that the degrees of repair of rat calvarial defect achieved with the drug-loaded nanofiber scaffolds were significantly better than those obtained with the blank materials; in particular, the dual-drug-loaded nanofiber scaffold manifested the best repair efficacy due to a synergistic effect of BMP-2 and DEX.

Schematic illustration of the fabrication of bone growth factors-in-polymer nanofiber device with coaxial electrospinning a and the nanofibers patches implanted in the dog leg bone defect b. It remains difficult to create clinically relevant 3D constructs beyond a relatively 2D mat. For bone tissue engineering, a large 3D scaffold may be required.

While new processing techniques have shown promise to increase the size and porosity of electrospun scaffolds. More work needs to be done to further help the architectural control. Having pores large enough for not only cell penetration, but also vascular in growth is imperative for a vascularized tissue such as bone.

Bone tissue itself represents a biological nanocomposite composed of organic predominantly collagen type I and inorganic nanocrystal-line HA components, with a hierarchical structure ranging from the microscale to the nanoscale. The inherent properties of nanocomposites, such as increased wettability, roughness, and surface area, can also promote biomaterial-driven bone regeneration through increased protein adsorption, nutrient exchange, and porosity relative to macroscale biomaterials.

Mehta et al. These findings indicated surface improvement and the applicability of this new nano-biomaterial for bone regenerative medicine. Nanocomposite scaffolds provide structural support for the cells, while changes to the nanoscale level of tissue hierarchy may have significant effects on cell-scaffold adhesion, integrin-triggered signaling pathways and cellular function; indeed, nanoscale features have been shown to have regulatory effects over multiple aspects of osteoblast and bone derived stem-cell behavior including adhesion, migration, proliferation, cell signaling, genetic expression, and stem cell fate.

Consequentially, biomaterial design has focused on the introduction of nanoscale elements that elicit directed cellular behavior while imparting structural and mechanical advantages to the bone construct to induce the formation of functional tissues. Current methodologies employed in the fabrication of nanocomposites include electrospinning and molecular self-assembly. The purpose of this article was to give a general description of studies of nanostructured materials for bone tissue engineering.

Nanophase ceramics, especially nano-HA, are popular bone substitutes, coatings and other filler materials due to their documented ability to promote mineralization. The nanometer grain sizes and high surface fraction of grain boundaries in nanoceramics increase osteoblast functions such as adhesion, proliferation, and differentiation.

Similar tendencies have been reported for other nanoceramics including alumina, zinc oxide, and titania; thus, providing evidence that, to some extent, it may not matter what implant chemistry is fabricated to have nanometer surface features to promote bone growth. However, this need further studies. For applications, synthetic and natural polymers, e. Nanoporous or nanofibrous polymer matrices can be fabricated via electrospinning, phase separation, particulate leaching, chemical etching, and 3D printing techniques.

Tissue Engineering and Regenerative Medicine: Achievements, Future, and Sustainability in Asia

For more stem cell books in this series, search Springer Link. It looks like you're using Internet Explorer 11 or older. This website works best with modern browsers such as the latest versions of Chrome, Firefox, Safari, and Edge. If you continue with this browser, you may see unexpected results. The purpose of this guide is to support the research needs of the Center. Also includes Clinical Genomics: Practical Applications in Adult Patient Care- an exploration of the molecular basis of common diseases by medical specialty, as well as images.

The mission of Nanomedicine: Nanotechnology, Biology, and Medicine Nanomedicine: NBM is to promote the emerging interdisciplinary field of nanomedicine. Nanomedicine: NBM is an international, peer-reviewed journal presenting novel, significant, and interdisciplinary theoretical and experimental results related to nanoscience and nanotechnology in the life and health sciences. Content includes basic, translational, and clinical research addressing diagnosis, treatment, monitoring, prediction, and prevention of diseases. Nanomedicine: NBM journal publishes articles on artificial cells, regenerative medicine, gene therapy, infectious disease, nanotechnology, nanobiotechnology, nanomedicine, stem cell and tissue engineering. Article types Selection of the appropriate article type is very important because requirements do differ. Length of communications should not exceed 1, words including body text, and figure legends , and the article should have no more than 5 figures.

Guide for Authors

It seems that you're in Germany. We have a dedicated site for Germany. Nanotechnology plays a key leading role in developing tools able to identify, measure, and study cellular events at the nanometric level as well as in contributing to the disclosure of unknown biological interactions and mechanisms, which opens the door for advances including nanodevices for diagnostic and therapy, drug delivery systems, and regenerative medicine. In Nanotechnology in Regenerative Medicine: Methods and Protocols , expert researchers in the field provide an overview of a very wide range of currently used technologies and methods that involve nanotechnology principles applicable to tissue regeneration.

Accurate and noninvasive stem cell tracking is one of the most important needs in regenerative medicine to determine both stem cell destinations and final differentiation fates, thus allowing a more detailed picture of the mechanisms involved in these therapies. Given the great importance and advances in the field of nanotechnology for stem cell imaging, currently, several nanoparticles have become standardized products and have been undergoing fast commercialization. This review has been intended to summarize the current use of different engineered nanoparticles in stem cell tracking for regenerative medicine purposes, in particular by detailing their main features and exploring their biosafety aspects, the first step for clinical application.

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Stem Cell Tracking with Nanoparticles for Regenerative Medicine Purposes: An Overview

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