Methylsulfonylmethane suppresses hepatic tumor development through activation of apoptosis.

AIM: To investigate the effect of methylsulfonylmethane (MSM), recently reported to have anti-cancer effects, in liver cancer cells and transgenic mice. METHODS: Three liver cancer cell lines, HepG2, Huh7-Mock and Huh7-H-ras (G12V), were used. Cell growth was measured by Cell Counting Kit-8 and soft agar assay. Western blot analysis was used to detect caspases, poly (ADP-ribose) polymerase (PARP), and B-cell lymphoma 2 (Bcl-2) expressions. For in vivo study, we administered MSM to H-ras (12V) transgenic mice for 3 mo. RESULTS: MSM decreased the growth of HepG2, Huh7-Mock and Huh7-H-ras (G12V) cells in a dose-dependent manner. That was correlated with significantly increased apoptosis and reduced cell numbers in MSM treated cells. Cleaved caspase-8, cleaved caspase-3 and cleaved PARP were remarkably increased in the liver cancer cells treated with 500 mmol/L of MSM; however, Bcl-2 was slightly decreased in 500 mmol/L. Liver tumor development was greatly inhibited in the H-ras (12V) transgenic mice treated with MSM, compared to control, by showing reduced tumor size and number. Cleaved PARP was significantly increased in non-tumor treated with MSM compared to control. CONCLUSION: Liver injury was also significantly attenuated in the mice treated with MSM. Taken together, all the results suggest that MSM has anti-cancer effects through inducing apoptosis in liver cancer.

Biofunctional porous anodized titanium implants for enhanced bone regeneration.

Efficient osseointegration is a key factor in dental implants to reduce the total time-course of therapy. Titanium implants with anodized surface gained much interest for their enhanced osseointegration. Anodized implant combined with bioactive drugs is an ideal candidate for enhance bone regeneration. Previously delivery of drugs from the metal implants has been attempted by utilizing a polymeric dip-coating method. However, the entire surface coating with polymer can diminish the advantageous surface roughness of anodized implants and cause contact inhibition between bone and implant surface. In this study, fibroblast growth factor-2 (FGF-2) loaded poly(lactide-co-glycolide) nanoparticles were partially coated on anodized Ti discs by an electrospray deposition. Nanoparticle coated anodized discs maintained their native porous structure and provided a sustained release of FGF-2 for more than 2 weeks with 40% initial burst. In vitro study confirmed the influence of polymeric nanoparticles and the release of growth factors from the Ti disc. Nanoparticle-coated groups significantly enhanced cell spreading and differentiation. For in vivo evaluation, the anodized titanium implants were applied to rabbit tibia model. The osseointegration was estimated by bone to implant contact of best three consecutive threads at the border of the implant. The mean osteointegration value of FGF-2 releasing implant groups (70.1%) was significantly higher than that of untreated implants (47.1%). We believe that the electrospray deposition technique is a particularly attractive approach for the coating of medical devices with porous surface to maintain their surface topography while allowing a sustained delivery of growth factors for bone regeneration. © 2013 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 102A: 3639-3648, 2014.

Suppression of thymus- and activation-regulated chemokine (TARC/CCL17) production by 3-O-ß-D-glucopyanosylspinasterol via blocking NF-κB and STAT1 signaling pathways in TNF-α and IFN-γ-induced HaCaT keratinocytes.

A phytosterol derivative, 3-O-ß-D-glucopyanosylspinasterol (spinasterol-Glc) isolated from leaves of Stewartia koreana was reported to inhibit LPS-induced cytokine production in macrophage cells. Thymus and activation regulated chemokine (TARC/CCL17) is produced in response to pro-inflammatory cytokines in keratinocytes, which is implicated in the development of inflammatory skin diseases. In present study, we investigated the effect of spinasterol-Glc on production of TARC/CCL17 induced by TNF-α and IFN-γ in human HaCaT keratinocytes. Spinasterol-Glc inhibited the mRNA and protein expression of TARC/CCL17 induced by TNF-α/IFN-γ in a dose-dependent manner. Inhibitors of c-Raf-1, p38 MAPK, and JAK2, suppressed the TNF-α/IFN-γ-induced production of TARC/CCL17, and phosphorylation of these signaling molecules were attenuated by spinasterol-Glc. The compound also inhibited phosphorylation of IKKα/ß and IκB-α, and reduced translocation of NF-κB to the nucleus. We demonstrated that spinasterol-Glc suppressed the NF-κB-driven and the GAS-driven expression of luciferase reporter gene induced by TNF-α and IFN-γ. In addition, spinasterol-Glc inhibited the DNA binding of NF-κB and STAT1 to its cognate binding site. These results suggest that spinasterol-Glc has effective inhibitory effects on production of TARC/CCL17 in keratinocytes via inhibition of NF-κB as well as STAT activation, and could be utilized for development of a potential therapeutic agent against skin inflammatory diseases.

Local BMP-7 release from a PLGA scaffolding-matrix for the repair of osteochondral defects in rabbits.

The use of tissue engineering to repair large osteochondral defects has been impeded by the limited regenerative capacity of cartilage. Herein, we describe the local release of bone morphogenetic protein 7 (BMP-7) to stimulate the bone marrow-derived progenitors to repair osteochondral defects. BMP-7-releasing poly(D,L-lactide-co-glycolide) (PLGA) matrix was specially designed to retain the dual-function of local BMP-7 release and progenitor-scaffolding with its defect-fitting architecture. To optimize the release kinetics during the repair period, BMP-7/PLGA film was cast on the surface of a cylindrical PLGA matrix. The matrix demonstrated a release profile of BMP-7 in a sustained manner over 28 days, maintaining its biological activity. The cylindrical PLGA matrices loaded with BMP-7 were implanted into the osteochondral defects (2 mm in diameter, 3 mm in depth) in rabbit knees. Histological observations revealed that neo-cartilage generation was completed in a well-integrated morphology with its surrounding normal cartilage and subchondral bone at 12 weeks post-implantation. Partial degradation of the PLGA matrix during the repair period guided neo-cartilage formation, which verified the effective scaffolding function of the matrix. Regenerated cartilage in BMP-7-treated defects stained positive for type II collagen and glycosaminoglycan (GAG). Adjacent BMP-7-untreated defects were also repaired with cartilage regeneration, suggesting the effect of local BMP-7 release in the synovial fluid. The BMP-7-unloaded PLGA matrix demonstrated guided cartilage regeneration to a certain extent, whereas the adjacent defects without the matrix revealed only fibrous tissue infiltration. These results indicated that a strategy employing the combined functions of local BMP-7 release and the cell scaffolding of a PLGA matrix might be a potential modality for osteochondral repair.

Controlled release of cell-permeable gene complex from poly(L-lactide) scaffold for enhanced stem cell tissue engineering.

The use of tissue engineering to deliver genes to stem cells has been impeded by low transfection efficiency of the inserted gene and poor retention at the target site. Herein, we describe the use of non-viral gene transfer by cell-permeable peptide (CPP) to increase the transfection efficiency. The combination of this technique with the use of a controlled release concept using a poly (l-lactide) scaffold allowed for prolonged uptake in stem cells. High transfection efficiency was obtained using a human-derived arginine-rich peptide denoted as Hph-1 (YARVRRRGPRR). The formation of complex between pDNA and Hph-1 was monitored using gel retardation tests to measure size and zeta potential. Complex formation was further assessed using a DNase I protection assay. A sustained gene delivery system was developed using a fibrous 3-D scaffold coated with pDNA/Hph-1 complexes. Transfection efficiency and the mean fluorescence intensity of human adipose-derived stem cells (hASCs) on the sustained delivery scaffold were compared to those of cells transfected via bolus delivery. Plasmid DNA completely bound Hph-1 at a negative-to-positive (N/P) charge ratio of 10. After complex formation, Hph-1 appeared to effectively protect pDNA against DNase I attack and exhibited cytotoxicity markedly lower than that of the pDNA/PEI complex. Plasmid DNA/Hph-1 complexes were released from the scaffolds over 14days and were successfully transfected hASCs seeded on the scaffolds. Flow cytometry revealed that the transfection efficiency in hASCs treated with pDNA/Hph-1 complex was approximately 5-fold higher than that in cells transfected using Lipofectamine. The sustained delivery system showed a significantly higher transfection efficiency and remained able to transfect cells for a longer period of time than bolus delivery. These results suggest that cell-scaffold-based tissue regeneration can be further improved by transduction concept using CPP and controlled release using polymeric scaffold.

Novel three-dimensional scaffolds of poly(L-lactic acid) microfibers using electrospinning and mechanical expansion: Fabrication and bone regeneration.

Poly(L-lactic acid) (PLLA) microfibrous scaffolds with three-dimensional (3D) structures were fabricated using an electrospinning technique with a subsequent mechanical expansion process. To achieve a 3D fibrous structure, the fusion at the contact points of the as-spun PLLA microfibers was avoided using an appropriate binary solvent system of methylene chloride and acetone. The solvent composition was optimized based on the solvent power, volatility, and viscosity (methylene chloride:acetone = 9:1 volume ratio). The final 3D structure of the electrospun scaffolds was obtained after mechanical expansion of the electrospun microfibrous mats. The pore sizes of the scaffolds were controlled by varying the degree of expansion of the nonbonded microfibrous mats, and they were in the range of several microns up to 400 µm. The 3D scaffolds were examined for their morphological properties and their potential use for the proliferation of osteoblasts. Generally recognized electrospun 2D nanofibrous membranes were also tested in order to compare the cell behaviors using different scaffold geometries. The 3D scaffolds demonstrated a high level of osteoblast proliferation (1.8-fold higher than nanofibrous membranes in a week). The osteoblasts actively penetrated the inside of the 3D scaffold and showed a spatial cell distribution, as confirmed by SEM and H&E staining, while a monolayer formed in the case of the 2D nanofibrous membranes with limited cell infiltration. In vivo results further showed that 3D electrospun microfibrous matrices were a favorable substrate for cell infiltration and bone formation after 2 and 4 weeks, using a rabbit calvarial defect model. In this study, the 3D microfibrous PLLA scaffolds fabricated using electrospinning techniques might be an innovative addition to tissue engineering applications.

Evaluation of the completeness of the nursing process for patients having gastrectomy using electronic nursing records.

The purpose of this study was to evaluate recording completeness of the nursing process. We compared nursing statements documented at the time when the Electronic Nursing Record (ENR) system, based on the ICNP, was implemented in 2004 with those documented in 2007. The ENRs for 35 gastrectomy patients in each year were selected for evaluation. The selected data were 11,822 nursing statements in 2004 and 27,870 in 2007. The results indicated a significant increase in the completeness of the nursing process in 2007. In addition, the number of nursing diagnosis increased by 5.1 times. The most contributing factor for this increase is assumed to be nurse education.