Preventive effects of epigallocatechin-3-O-gallate against replicative senescence associated with p53 acetylation in human dermal fibroblasts

Considering the various pharmacological activities of epigallocatechin-3-O-gallate (EGCG) including anticancer, and anti-inflammatory, antidiabetic, and so forth, relatively less attention has been paid to the antiaging effect of EGCG on primary cells. In this study, the preventive effects of EGCG against serial passage-induced senescence were investigated in primary cells including rat vascular smooth muscle cells (RVSMCs), human dermal fibroblasts (HDFs), and human articular chondrocytes (HACs). The involvement of Sirt1 and acetylated p53 was examined as an underlying mechanism for the senescence preventive activity of EGCG in HDFs. All cells were employed with the initial passage number (PN) between 3 and 7. For inducing senescence, the cells were serially passaged at the predetermined times and intervals in the absence or presence of EGCG (50 or 100 μM). Serial passage-induced senescence in RVSMCs and HACs was able to be significantly prevented at 50 μM EGCG, while in HDFs, 100 μM EGCG could significantly prevent senescence and recover their cell cycle progression close to the normal level. Furthermore, EGCG was found to prevent serial passage- and H(2)O(2)-induced senescence in HDFs by suppressing p53 acetylation, but the Sirt1 activity was unaffected. In addition, proliferating HDFs showed similar cellular uptake of FITC-conjugated EGCG into the cytoplasm with their senescent counterparts but different nuclear translocation of it from them, which would partly account for the differential responses to EGCG in proliferating versus senescent cells. Taking these results into consideration, it is suggested that EGCG may be exploited to craft strategies for the development of an antiaging or age-delaying agent.ope

Effect of electron beam irradiation on load-displacement responses of UHMWPE

Ultra-High Molecular Weight Polyethylene (UHMWPE) has been used successfully for articulating surfaces of joint endoprostheses, i.e. cup linear on acetabular component of total hip arthroplasty, in orthopaedics for over three decades. Although numerous attempts have been performed to characterize and to improve the polymeric material, i.e. to reduce the molecular degradation due to in vivo oxidation, the effect of electron beam irradiation on loaddisplacement characteristics of UHMWPE has not been reported clearly. The aim of the current study is to quantify the effect of the electron beam irradiation with alteration of the amount of irradiation on the mechanical characteristics of the polymeric material. In this work, monotonic uniaxial and small punch tests were conducted to examine the relative capabilities of four samples irradiated with electron beam at 0, 50, 100, and 150 kGy, based on a prediction of mechanical behavior of the UHMWPE materials. Load-displacement behaviors of UHMWPE were not changed significantly by electron beam irradiation less than 150 kGy although the stiffness of irradiated UHMWPE decreased gradually. Conclusively, changes of stiffness and deflection by irradiation were not significant. However, load characteristics were significantly changed by irradiation.ope

Recent Advances in ROS-Responsive Cell Sheet Techniques for Tissue Engineering

Cell sheet engineering has evolved rapidly in recent years as a new approach for cell-based therapy. Cell sheet harvest technology is important for producing viable, transplantable cell sheets and applying them to tissue engineering. To date, most cell sheet studies use thermo-responsive systems to detach cell sheets. However, other approaches have been reported. This review provides the progress in cell sheet detachment techniques, particularly reactive oxygen species (ROS)-responsive strategies. Therefore, we present a comprehensive introduction to ROS, their application in regenerative medicine, and considerations on how to use ROS in cell detachment. The review also discusses current limitations and challenges for clarifying the mechanism of the ROS-responsive cell sheet detachment.ope

Preconditioning process for dermal tissue decellularization using electroporation with sonication

Decellularization to produce bioscaffolds composed of the extracellular matrix (ECM) uses enzymatic, chemical and physical methods to remove antigens and cellular components from tissues. Effective decellularization methods depend on the characteristics of tissues, and in particular, tissues with dense, complex structure and abundant lipid content are difficult to completely decellularize. Our study enables future research on the development of methods and treatments for fabricating bioscaffolds via decellularization of complex and rigid skin tissues, which are not commonly considered for decellularization to date as their structural and functional characteristics could not be preserved after severe decellularization. In this study, decellularization of human dermal tissue was done by a combination of both chemical (0.05% trypsin-EDTA, 2% SDS and 1% Triton X-100) and physical methods (electroporation and sonication). After decellularization, the content of DNA remaining in the tissue was quantitatively confirmed, and the structural change of the tissue and the retention and distribution of ECM components were evaluated through histological and histochemical analysis, respectively. Conditions of the chemical pretreatment that increase the efficiency of physical stimulation as well as decellularization, and conditions for electroporation and sonication without the use of detergents, unlike the methods performed in previous studies, were established to enable the complete decellularization of the skin tissue. The combinatorial decellularization treatment formed micropores in the lipid bilayers of the skin tissues while removing all cell and cellular residues without affecting the ECM properties. Therefore, this procedure can be widely used to fabricate bioscaffolds by decellularizing biological tissues with dense and complex structures.ope

Enhanced Neural Cell Adhesion and Neurite Outgrowth on Graphene-Based Biomimetic Substrates

Neural cell adhesion and neurite outgrowth were examined on graphene-based biomimetic substrates. The biocompatibility of carbon nanomaterials such as graphene and carbon nanotubes (CNTs), that is, single-walled and multiwalled CNTs, against pheochromocytoma-derived PC-12 neural cells was also evaluated by quantifying metabolic activity (with WST-8 assay), intracellular oxidative stress (with ROS assay), and membrane integrity (with LDH assay). Graphene films were grown by using chemical vapor deposition and were then coated onto glass coverslips by using the scooping method. Graphene sheets were patterned on SiO2/Si substrates by using photolithography and were then covered with serum for a neural cell culture. Both types of CNTs induced significant dose-dependent decreases in the viability of PC-12 cells, whereas graphene exerted adverse effects on the neural cells just at over 62.5 ppm. This result implies that graphene and CNTs, even though they were the same carbon-based nanomaterials, show differential influences on neural cells. Furthermore, graphene-coated or graphene-patterned substrates were shown to substantially enhance the adhesion and neurite outgrowth of PC-12 cells. These results suggest that graphene-based substrates as biomimetic cues have good biocompatibility as well as a unique surface property that can enhance the neural cells, which would open up enormous opportunities in neural regeneration and nanomedicine.ope

Ethyl-3,4-dihydroxybenzoate with a dual function of induction of osteogenic differentiation and inhibition of osteoclast differentiation for bone tissue engineering.

The current approach in biomaterial design of bone implants is to induce in situ regeneration of bone tissue, thus improving integration of the implants and reducing their failure. Therefore, ethyl-3,4-dihydroxybenzoate (EDHB), which stimulates differentiation of osteoblasts and the resultant bone formation, should be studied. In this study, the osteoinductive ability of EDHB in preosteoblasts and human mesenchymal stem cells was examined. EDHB for future use in bone tissue engineering was evaluated by examination of early markers of differentiation (such as alkaline phosphatase [ALP] activity and collagen type I expression) and late markers of osteoblast differentiation (bone nodule formation). As bone remodeling and implant osteointegration depend not only on osteoblast response but also on interaction of the biomaterial with bone-resorbing osteoclasts, differentiation of osteoclasts in response to the compounds was also observed. For in vivo study, alginate gel comprised of EDHB and cells was transplanted into the back subcutis of mice. Our results show that EDHB might have beneficial effects through regulation of both osteoblast and osteoclast differentiation. Therefore, we suggest that EDHB could be a strong candidate for dual regulation to increase osteoblast differentiation and decrease osteoclast differentiation.ope

An effective method to generate controllable levels of ROS for the enhancement of HUVEC proliferation using a chlorin e6-immobilized PET film as a photo-functional biomaterial

Reactive oxygen species (ROS) are byproducts of cellular metabolism; they play a significant role as secondary messengers in cell signaling. In cells, high concentrations of ROS induce apoptosis, senescence, and contact inhibition, while low concentrations of ROS result in angiogenesis, proliferation, and cytoskeleton remodeling. Thus, controlling ROS generation is an important factor in cell biology. We designed a chlorin e6 (Ce6)-immobilized polyethylene terephthalate (PET) film (Ce6-PET) to produce extracellular ROS under red-light irradiation. The application of Ce6-PET films can regulate the generation of ROS by altering the intensity of light-emitting diode sources. We confirmed that the Ce6-PET film could effectively promote cell growth under irradiation at 500 μW/cm2 for 30 min in human umbilical vein endothelial cells. We also found that the Ce6-PET film is more efficient in generating ROS than a Ce6-incorporated polyurethane film under the same conditions. Ce6-PET fabrication shows promise for improving the localized delivery of extracellular ROS and regulating ROS formation through the optimization of irradiation intensity.ope

The Control of Pyrogenicity and Sterility for The Biomaterials in Life Cycle Perspective

The proper sterilization methods for biomaterials as eradicating harmful microorganisms is very important safety element to prevent infections of patients before transplanting biomaterials. The sterilization process to eradicate microorganisms, which is a special process in a sense that the degree of sterilization cannot be fully verified, requires validation methods. Therefore, a comprehensive quality system is essential throughout the entire process, such as prevention of microorganism contamination, sterilization process and maintenance of sterility for the control of pyrogenicity and sterility. This paper describes (1) a method to control pyrogenicity of medical devices that contact with blood, such as artificial blood vessels, blood dialyzer and stent, provides (2) an understanding of the concept of sterilization validation as a means to demonstrate sterility assurance level (SAL) and reviews (3) the application techniques of sterilization validation method to achieve SAL of invasive medical devices in the perspective of life cycle, focusing on the half cycle method.ope

Selective sterilization of Vibro parahaemolyticus from a bacterial mixture by low-amperage electric current

The objective of this study was to investigate the possibility of using low-amperage electrical treatment (LAET) as a selective bacteriocide. Mixtures containing Escherichia coli, Staphylococcus aureus, and Vibrio parahaemolyticus were treated with different electric current intensities and for different times. The results showed that at 263 mA, treating bacteria for 100 ms eliminated all V. parahaemolyticus colonies. Although LAET reduced the populations of the three microorganisms, V. parahaemolyticus was more injured by LAET than S. aureus and E. coli when treated at the same processing conditions.ope

Inactivation of Listeria monocytogenes in brine and saline by alternating high-voltage pulsed current

The inactivating efficiency of alternating high-voltage pulsed (AHVP) current was investigated in brine (20 w/v% NaCl) and saline (0.9 w/v% NaCl) inoculated with 1x 10(7) cells/ml of Listeria monocytogenes. AHVP current at 12 V with 1 pulse completely inactivated L. monocytogenes in brine within 3 ms, while the bacteria in saline were fully inactivated by 10-pulsed electric treatment at 12 V within the same time. Electron microscopic observation demonstrated substantial structural damage of electrically treated L. monocytogenes in brine. These results suggest that AHVP treatment would be effective for the rapid and complete inactivation of L. monocytogenes in brine or saline solution.ope