Toward Biomimetic Scaffolds for Tissue Engineering: 3D Printing Techniques in Regenerative Medicine.

Three-dimensional (3D) printing technology allows fabricating complex and precise structures by stacking materials layer by layer. The fabrication method has a strong potential in the regenerative medicine field to produce customizable and defect-fillable scaffolds for tissue regeneration. Plus, biocompatible materials, bioactive molecules, and cells can be printed together or separately to enhance scaffolds, which can save patients who suffer from shortage of transplantable organs. There are various 3D printing techniques that depend on the types of materials, or inks, used. Here, different types of organs (bone, cartilage, heart valve, liver, and skin) that are aided by 3D printed scaffolds and printing methods that are applied in the biomedical fields are reviewed.

Skin Regeneration with a Scaffold of Predefined Shape and Bioactive Peptide Hydrogels.

We developed a highly elastic customized scaffold for soft tissue regeneration and combined them with bioactive hydrogels with stem cell-inducing ability. This was done to mimic mechanical properties of native soft tissues and improve the viability of transplanted cells as well as efficiency of tissue regeneration. The proposed study was aimed at evaluating various characteristics of scaffolds and investigating their tissue-regenerating ability. Finger-shaped porous scaffolds were successfully fabricated by an indirect 3D printing of poly (L-lactide-co-ɛ-caprolactone) (PLCL), which provides high elasticity for soft tissue engineering. In addition, a self-assembling peptide hydrogel coupled with substance P (RARADADARARADADA/RARADADARARADADA-substance P, RADA16/RADA16-SP) was used to accelerate angiogenesis and recruit intrinsic mesenchymal stem cells (MSCs). This study included three kinds of groups: Group I = PLCL scaffold with human dermal fibroblasts (HDFs) (P+C), Group II = PLCL scaffold with HDFs and RADA16 (P+C+R), and Group III = PLCL scaffold with HDFs, RADA16, and RADA16-SP (P+C+R+S). The samples were implanted into immunodeficient mice subcutaneously and harvested at 1 and 4 weeks. Tissue regeneration was evaluated by histological analysis with hematoxylin and eosin (H&E) and Masson's trichrome (MT) staining. The images showed that a large number of cells were recruited into the scaffolds, and collagen was deposited in the constructs of the P+C+R+S group. Additionally, recruitment of MSCs, angiogenesis, and collagen were observed by immunofluorescence staining. The results show that the P+C+R+S group had more type I and type III collagen, which are formed in soft tissues, and were deposited on the scaffold compared with the other groups. Moreover, more blood vessels and MSCs were induced in the P+C+R+S group than in those of the P + C and P+C+R groups. Consequently, the results suggest that the construct of the customized porous PLCL scaffold and RADA16/RADA16-SP hydrogel could be a good treatment modality to treat skin defects.

Human steroid sulfatase induces Wnt/ß-catenin signaling and epithelial-mesenchymal transition by upregulating Twist1 and HIF-1α in human prostate and cervical cancer cells.

Steroid sulfatase (STS) catalyzes the hydrolysis of estrone sulfate and dehydroepiandrosterone sulfate (DHEAS) to their unconjugated biologically active forms. Although STS is considered a therapeutic target for estrogen-dependent diseases, the cellular functions of STS remain unclear. We found that STS induces Wnt/ß-catenin s Delete ignaling in PC-3 and HeLa cells. STS increases levels of ß-catenin, phospho-ß-catenin, and phospho-GSK3ß. Enhanced translocation of ß-catenin to the nucleus by STS might activate transcription of target genes such as cyclin D1, c-myc, and MMP-7. STS knockdown by siRNA resulted in downregulation of Wnt/ß-catenin signaling. ß-Catenin/TCF-mediated transcription was also enhanced by STS. STS induced an epithelial-mesenchymal transition (EMT) as it reduced the levels of E-cadherin, whereas levels of mesenchymal markers such as N-cadherin and vimentin were enhanced. We found that STS induced Twist1 expression through HIFα activation as HIF-1α knockdown significantly blocks the ability of STS to induce Twist1 transcription. Furthermore, DHEA, but not DHEAS is capable of inducing Twist1. Treatment with a STS inhibitor prevented STS-mediated Wnt/ß-catenin signaling and Twist1 expression. Interestingly, cancer cell migration, invasion, and MMPs expression induced by STS were also inhibited by a STS inhibitor. Taken together, these results suggest that STS induces Wnt/ß-catenin signaling and EMT by upregulating Twist1 and HIF-1α. The ability of STS to induce the Wnt/ß-catenin signaling and EMT has profound implications on estrogen-mediated carcinogenesis in human cancer cells.

Fabrication of porous chitosan-polyvinyl pyrrolidone scaffolds from a quaternary system via phase separation.

Three-dimensional porous chitosan-polyvinyl pyrrolidone (PVP) scaffolds were fabricated for tissue engineering applications via liquid-liquid or liquid-solid phase separation. A mixture of an acidic aqueous solution with butanol as a non-solvent and a chitosan-PVP quaternary system were freeze-dried. We then studied the homogenous open pore structure and the minute pore distribution in order to improve the mass transfer and cell seeding efficiency while also obtaining the optimal ratio of PVP to provide high interconnectivity and to improve the open-pore structure. The properties of the porous chitosan-PVP scaffolds - including the microstructure, chemical release, water absorption properties, and cell proliferation tests were studied - and the results were compared against those obtained from conventional scaffolds. chitosan-PVP scaffolds with a porosity of over 70% were obtained, and the pore morphology on the surface and within the porous scaffolds showed the presence of homogenous open pores with excellent interconnectivity. As the PVP content increased, main pores (50-100 µm) and minute pores (4-10 µm) could be clearly observed. Also, the porous scaffold showed an improved efficiency for cell adhesion after the cells were cultured for 4 h. After 72 h, the cultured cells presented an increase in the cell proliferation and on the porous scaffolds. These results strongly suggest that the porous chitosan-PVP scaffolds can be widely used in tissue engineering, including for biopatches and artificial skin applications.

Preparation and characterization of Ag nanoparticle-embedded blank and ligand-anchored silica gels.

Ag nanoparticles, used for halogen (especially iodine) adsorption and an evaluation of halogen behavior, were embedded in synthesized inorganic-organic hybrid gels. In particular, an irradiation method using an electron beam plays a part in introducing Ag nanoparticles to the organofunctionalized silica gels from AgNO3 solutions in a simple way at atmospheric pressure and room temperature. For preparation of the Ag nanoparticle-embedded inorganic-organic hybrid gels, ligands of ethylenediamine (NH2CH2CH2NH-, TMSen) and mercapto (HS-) functionalized three-dimensional porous SiO2 sol-gels were first synthesized through hydrolysis and condensation reactions, and Ag nanoparticles were then embedded into the ethylenediamine- and mercapto-anchored silica gels each, through electron-beam irradiation. The addition of ligands yielded larger average pore sizes than the absence of any ligand. Moreover, the ethylenediamine ligand led to looser structures and better access of the Ag nanoparticles to the ethylenediamine-anchored gel. As a result, more Ag nanoparticles were introduced into the ethylenediamine-anchored gel. The preparation and characterization of Ag nanoparticle-embedded blank and ligand-anchored silica gels are discussed in detail.

Induction of steroid sulfatase expression in PC-3 human prostate cancer cells by insulin-like growth factor II.

Human steroid sulfatase (STS) plays an important role in regulating the formation of biologically active estrogens and may be a promising target for treating estrogen-mediated carcinogenesis. The molecular mechanism of STS gene expression, however, is still not clear. Growth factors are known to increase STS activity but the changes in STS expression have not been completely understood. To determine whether insulin-like growth factor (IGF)-II can induce STS gene expression, the effects of IGF-II on STS expression were studied in PC-3 human prostate cancer cells. RT-PCR and Western blot analysis showed that IGF-II treatment significantly increased the expression of STS mRNA and protein in concentration- and time-dependent manners. To understand the signaling pathway by which IGF-II induces STS gene expression, the effects of specific PI3-kinase/Akt and NF-κB inhibitors were determined. When the cells were treated with IGF-II and PI3-kinase/Akt inhibitors, such as LY294002, wortmannin, or Akt inhibitor IV, STS expression induced by IGF-II was significantly blocked. Moreover, we found that NF-κB inhibitors, such as MG-132, bortezomib, Bay 11-7082 or Nemo binding domain (NBD) binding peptide, also strongly prevented IGF-II from inducing STS gene expression. We assessed whether IGF-II activates STS promoter activity using transient transfection with a luciferase reporter. IGF-II significantly stimulated STS reporter activity. Furthermore, IGF-II induced expression of 17ß-hydroxysteroid dehydrogenase (HSD) 1 and 3, whereas it reduced estrone sulfotransferase (EST) gene expression, causing enhanced estrone and ß-estradiol production. Taken together, these results strongly suggest that IGF-II induces STS expression via a PI3-kinase/Akt-NF-κB signaling pathway in PC-3 cells and may induce estrogen production and estrogen-mediated carcinogenesis.

Radiolytic production of Ag-containing nanocomposite colloids in presence of lithium ions.

This research is motivated to prepare homogeneous and less aggregated nanostructured composites for potential scintillators. Poly ethylene glycol (PEG)-protected approximately 5 nm and approximately 29 nm sized Ag nanoparticles and Ag-Zn nanocomposites were relatively prepared by electron-beam irradiation on Ag+ and Zn2+ solutions with the aids of stabilizers and 6Li+ at room temperature under atmospheric pressure. Especially the 6Li+, which was used for a neutron absorption purpose, played a part the Ag-Zn nanocolloids to less aggregate in the aqueous phase by making partial complexes with stabilizers containing Ag-Zn. To be a potential scintillator, the Ag-Zn nanocomposites have to show an optical response to radiation. Therefore, optical luminescence, which resembles the concept of detecting light without the requirement of a neutron absorbent (convertor) for a neutron scitillator, of the nanocomposites was tested.

Adsorption behavior of Eu(III) on partially Fe(III)- or Ti(IV)-coated silica.

The adsorption behavior of Eu(III) onto silica surface, which was partially coated with Fe(III) or Ti(IV), was investigated to determine Fe(III) or Ti(IV) effects on the surface reaction of lanthanides on mineral surfaces in groundwater. Compared with a parallel uncoated silica, the Fe(III)-coated silica did not enhance the adsorption of Eu(III). However, enhanced adsorption of Eu(III) on the Ti(IV)-coated silica was observed by increasing the amount of Ti(IV) on the silica surface.

Bacterial Lipopolysaccharides Induce Steroid Sulfatase Expression and Cell Migration through IL-6 Pathway in Human Prostate Cancer Cells.

Steroid sulfatase (STS) is responsiblefor the conversion of estrone sulfate to estrone that can stimulate growth in endocrine-dependent tumors such as prostate cancer. Although STS is considered as a therapeutic target for the estrogen-dependent diseases, cellular function of STS are still not clear. Previously, we found that tumor necrosis factor (TNF)-α significantly enhances steroid sulfatase expression in PC-3 human prostate cancer cells through PI3K/Akt-dependent pathways. Here, we studied whether bacterial lipopolysaccharides (LPS) which are known to induce TNF-α may increase STS expression. Treatment with LPS in PC-3 cells induced STS mRNA and protein in concentration- and time-dependent manners. Using luciferase reporter assay, we found that LPS enhanced STS promoter activity. Moreover, STS expression induced by LPS increased PC-3 tumor cell migration determined by wound healing assay. We investigated that LPS induced IL-6 expression and IL-6 increased STS expression. Taken together, these data strongly suggest that LPS induces STS expression through IL-6 pathway in human prostate cancer cells.

Influence of zinc precursors and surfactants on the preparation of Ag-containing materials by electron beam irradiation.

Ag-Zn bimetallic nanocomposites or Ag2S precipitates were produced by irradiating a mixture of AgNO3 and Zn(NO3)2, and another mixture of AgNO3 and ZnS with an electron beam in an aqueous phase. Surfactants of poly vinyl alcohol (PVA) and poly ethylene glycol (PEG) induced a slightly different formation of particles during this process.

Temperature dependence of fluorescence for EuCl3 in LiCl-KCl eutectic melt.

The fluorescence of EuCl(3) in LiCl-KCl eutectic melt according to temperature changes was investigated, and the spontaneous partial reduction of Eu(3+) to Eu(2+) at high temperature was confirmed by the fluorescence results. The fluorescence decreases when the temperature increases, and this was examined in detail. The studies of fluorescence provided information regarding the chemical and physical behavior of europium ions in the molten salt according to the temperature changes. It is applicable for monitoring species and concentrations and estimating the approximate chemical structure of the ions in molten salts.

Potential application of fabricated sulfide-based scintillation materials for radiation detection.

In our laboratories, we have produced ZnS(Ag)/6Li sol-gel scintillation materials which produce an excellent light output with an alpha radiation (compared to commercial high temperature lithiated glass; KG-2 and a plastic scintillator; BC-400). However, when tested with a neutron radiation, the opacity of the ZnS(Ag)/6Li sol-gel scintillation materials, which were composed of a homogeneous micron-sized ZnS(Ag), prevented a clear neutron energy peak formation, thus making it difficult to set a threshold for neutron-gamma discrimination. In an effort to increase the transparency of the scintillation materials and to develop new technologies to fabricate sulfide-based scintillation materials for neutron detection, we turned to the methods of a chemical bath deposition (CBD) and a nano-particle synthesis for possible solutions.

Classification of materials for explosives from prompt gamma spectra by using principal component analysis.

Data from the elemental composition-ratios and experimental prompt gamma spectra of samples were used to develop suitable discriminant classes for suspect samples. The collected data and gamma spectra were applied to principal component analysis (PCA) to discriminate explosives from non-explosive materials.

Neutron scintillators of transparent silica xerogel monolith via a sealed container system and pi-pi interactions.

Transparent crack-free lithiated sol-gel scintillating monoliths were developed by taking advantage of a sealed container system for a syneresis and the pi-pi interactions between sol-gel components and organic fluors to yield a better homogeneity and scintillating efficiency. The transparency of the resulting materials indicates that the new scintillating material composites are mesoscopically dispersed. The silica monolith can be prepared without cladding the monolith with an engineering plastic such as a poly(ether ether ketone) (PEEK) or a liquid mounting medium. A successful detection of neutron particles by using these lithiated scintillating monoliths was demonstrated.

Synthesis and characterization of periodic mesoporous organosilicas as anion exchange resins for perrhenate adsorption.

A new methodology to immobilize ionic liquids through the use of a bridged silsesquioxane N-(3-triethoxysilylpropyl), N(3)-(3-trimethoxysilylpropyl-4,5-dihydroimidazolium iodide that incorporates an ionic functionality for the assembly of novel periodic mesoporous organosilica (PMO) materials has been developed. The resulting PMO materials were investigated for use as novel anion exchange resins for the separation of perrhenate anions in aqueous solution. As compared with cetyltrimethylammonium chloride, 1-hexadecane-3-methylimidazolium bromide has been demonstrated to be a more efficient surfactant template for the generation of mesopores and surface areas for such PMO materials.

Facile, alternative synthesis of lanthanum phosphate nanocrystals by ultrasonication.

Highly luminescent, rhabdophane (Ce(0.33)La(0.66))PO4.nH2O nanorods and nanoparticles were prepared in aqueous solutions by ultrasonication, at pH 1 and pH 12, respectively. Both nanorods (5 to 9 nm wide and several tens to several hundreds nm long) and nanoparticles (elongated, connected 5 nm particles) were as small and as uniform as products obtained from methods that utilize complexing agents or surfactants, only with no complexing agent. This method of synthesis by ultrasonication is a fast and simple method and it is expected to be applicable for the synthesis of other nanocrystalline lanthanide phosphates.

Mechanistic investigation of hydrolysis reactions of dithioacetal derivatives grafted on silica gels.

Silica gels believed to be grafted with dithioacetal derivatives were recently used for Hg(II) extraction, and were found to selectively remove 94-100% of Hg(2+) ions from metal ion mixtures. The current studies with one derivative suggest that the functional group in the Hg(II) removal is the mercapto (SH) ligand. The dithioacetal group in (ClCOCH(2)S)(2)CHPh (2) was hydrolyzed during its reaction with amine (O)(3)Si(CH(2))(3)NH(2) (3) grafted on silica gel to give the mercapto ligand (O)(3)Si(CH(2))(3)NHCOCH(2)SH (4). The silica gel grafted with the mercapto ligand 4 selectively removed Hg(2+) ions with reported high capacities.