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1.
Biomaterials ; 272: 120785, 2021 05.
Article in English | MEDLINE | ID: mdl-33819813

ABSTRACT

This study aimed to fabricate a multi-layered biphasic calcium phosphate (BCP) platform for programmed bone morphogenetic protein-2 (BMP-2) release, which means to block the initial burst release and promote releasing during the differentiation phase of osteogenic cells. And it is to confirm in vivo whether this platform has osteogenic inductivity even when extremely low doses of BMP-2 are loaded compared to the conventional soaking method. Our strategy consisted of preparing a multilayer coating on BCP to minimize the contact between BMP-2 and BCP and allow the loading of BMP-2. The multilayer, which is surface-modified on BCP, is composed of an organosilicate and a natural polymer-based layer-by-layer (LbL) film. We applied (3-Aminopropyl)triethoxysilane (APTES) as an organosilicate was used for amine-functionalized BCP and (collagen/heparin)5 film was used to delay and sustain BMP-2 release. The coated multilayer not only reduced the initial burst release by more than 50% but also loaded more BMP-2. For in vivo experiment, histomorphometric analysis, it was observed that the BCP platform loaded with extremely low concentration BMP-2 (0.01 mg/ml) induced a significantly larger amount of new bones at 8 weeks compared to the conventional soaking method in the rabbit calvarium onlay graft model.


Subject(s)
Bone Morphogenetic Protein 2 , Tissue Scaffolds , Animals , Bone Regeneration , Calcium Phosphates , Hydroxyapatites , Osteogenesis , Rabbits
2.
Sci Total Environ ; 752: 142242, 2021 Jan 15.
Article in English | MEDLINE | ID: mdl-33207500

ABSTRACT

Microplastics which are gradually and randomly decompose into small fragment by exposure of physical and biological external stress are emerging as a significant threat to the all the environments. Here, we have demonstrated the in vitro toxicity of microplastics of two different shapes. To minimize the chemical effect, polyethylene (PE), was used. PE microplastics with two different shapes were prepared, high-density PE microbeads and irregularly ground low-density PE from bulk pellets. It is hypothesized that morphological characteristics and concentration of PE microplastics could affect cellular viability, immunity, and lysis. To quantify the randomness of the microplastic shape, the edge patterns of the generated PE microplastics were converted into numerical values and analyzed using a statistical method. A 10-fold difference in curvature value was observed between microbeads and ground microfragments. To correlate shape differences to toxicology, cells were exposed to PE microplastics on the demand of toxicology studies. We found that the higher concentration and rough structure were associated with the toxicity of plastics toward cells, pro-inflammatory cytokine release, and hemolysis, even though PE is buoyant onto medium. The PE microbeads did not exhibit severe cytotoxicity at any of the tested concentrations, but induced immune and hemolysis responses at high concentrations. When comparing the toxicity of different shapes of PE microplastics, we confirmed by statistical analysis that irregular-shape plastics with sharp edges and higher curvature differences may adversely affect cells, further having possibility to human toxicity in real environment.


Subject(s)
Polyethylene , Water Pollutants, Chemical , Environmental Monitoring , Microplastics , Plastics/toxicity , Polyethylene/analysis , Polyethylene/toxicity , Water Pollutants, Chemical/analysis , Water Pollutants, Chemical/toxicity
3.
Sci Rep ; 10(1): 7391, 2020 04 30.
Article in English | MEDLINE | ID: mdl-32355311

ABSTRACT

Environmental pollution arising from plastic waste is a major global concern. Plastic macroparticles, microparticles, and nanoparticles have the potential to affect marine ecosystems and human health. It is generally accepted that microplastic particles are not harmful or at best minimal to human health. However direct contact with microplastic particles may have possible adverse effect in cellular level. Primary polystyrene (PS) particles were the focus of this study, and we investigated the potential impacts of these microplastics on human health at the cellular level. We determined that PS particles were potential immune stimulants that induced cytokine and chemokine production in a size-dependent and concentration-dependent manner.

4.
Sci Total Environ ; 684: 657-669, 2019 Sep 20.
Article in English | MEDLINE | ID: mdl-31158627

ABSTRACT

Environmental pollution caused by plastic waste is a growing global problem. Discarded plastic products and debris (microplastic particles) in the oceans detrimentally affect marine ecosystems and may impact human. Humans are exposed to plastic debris via the consumption of seafood and drinking water, contact with food packaging, or inhalation of particles. The accumulation of microplastic particles in humans has potential health risks such as cytotoxicity, hypersensitivity, unwanted immune response, and acute response like hemolysis. We investigated the cellular responses of secondary polypropylene microplastics (PP particles) of approximately ~20 µm and 25-200 µm in different condition and size to normal cells, immune cells, blood cells, and murine immune cells by cytokine analysis, ROS assay, polarization assay and proliferation assay. We found that PP particles showed low cytotoxicity effect in size and concentration manner, however, a high concentration, small sized, DMSO method of PP particles stimulated the immune system and enhanced potential hypersensitivity to PP particles via an increase in the levels of cytokines and histamines in PBMCs, Raw 264.7 and HMC-1 cells.


Subject(s)
Leukocytes/drug effects , Particulate Matter/toxicity , Polypropylenes/toxicity , Water Pollutants, Chemical/toxicity , Animals , Cell Line , Humans , Mice , Particle Size , RAW 264.7 Cells
5.
Biotechnol Bioeng ; 115(5): 1331-1339, 2018 05.
Article in English | MEDLINE | ID: mdl-29337363

ABSTRACT

Animal cell culture technology for therapeutic protein production has shown significant improvement over the last few decades. Chinese hamster ovary (CHO) cells have been widely adapted for the production of biopharmaceutical drugs. In the biopharmaceutical industry, it is crucial to develop cell culture media and culturing conditions to achieve the highest productivity and quality. However, CHO cells are significantly affected by apoptosis in the bioreactors, resulting in a substantial decrease in product quantity and quality. Thus, to overcome the obstacle of apoptosis in CHO cell culture, it is critical to develop a novel method that does not have minimal concern of safety or cost. Herein, we showed for the first time that exosomes, which are nano-sized extracellular vesicles, derived from CHO cells inhibited apoptosis in CHO cell culture when supplemented to the culture medium. Flow cytometric and microscopic analyses revealed that substantial amounts of exosomes were delivered to CHO cells. Higher cell viability after staurosporine treatment was observed by exosome supplementation (67.3%) as compared to control (41.1%). Furthermore, exosomes prevented the mitochondrial membrane potential loss and caspase-3 activation, meaning that the exosomes enhanced cellular activities under pro-apoptotic condition. As the exosomes supplements are derived from CHO cells themselves, it is not only beneficial for the biopharmaceutical productivity of CHO cell culture to inhibit apoptosis, but also from a regulatory standpoint to diminish any safety concerns. Thus, we conclude that the method developed in this research may contribute to the biopharmaceutical industry where minimizing apoptosis in CHO cell culture is beneficial.


Subject(s)
Apoptosis , CHO Cells/drug effects , CHO Cells/physiology , Cell Culture Techniques/methods , Exosomes/metabolism , Animals , Cell Survival , Cricetulus , Culture Media/chemistry , Flow Cytometry , Microscopy
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