The Hollow Porous Sphere Cell Carrier for the Dynamic Three-Dimensional Cell Culture.

Large-scale mammalian cell culture is essential in cell therapy, vaccine production, and the manufacturing of therapeutic protein drugs. Due to the adherent growth characteristic of most mammalian cell types, the combination of cell carrier and bioreactor is a common choice in large-scale mammalian cell culture. Current cell carriers developed by polymer crosslinking, lithography, or emulsion drops are unable to obtain a structure with uniformed porous structure and porous interior design, which results in an inhomogeneous culture condition for cells and therefore cannot ensure an optimal dynamic culture condition for cell proliferation, matrix production, and cell differentiation. In addition, the fluidic shear stress (a standard mechanical stimulation in bioreactor culture) and inner-carrier velocity (to ensure nutrient transport and waste exchange), which influence cell viability and growth, are not well-controlled/analyzed due to an irregular porous structure with these traditionally synthesized cell carriers. To solve these problems, we designed four types of hollow porous spheres (HPS, 1.0 cm diameter) with different porous structures. To investigate the impacts of porous structure on surface shear stress and inner velocity, computational fluid dynamics (CFD) simulations were conducted to analyze the liquid flow behavior in HPSs, based on which an optimal structure with minimal surface shear stress and best inner velocity was obtained and fabricated using fused deposition modeling three-dimensional (3D) printing technology. Inspired by the industrial large-scale culture system, a novel 3D dynamic culture system was then established using HPSs to seed the cells, which were then placed in a mini bioreactor on a tube roller. CFD analysis showed that under 0.1 m/s water flow, the shear stress at most surface areas from four HPSs was lower than 20 dynes/cm2, which suggests that the HPSs should provide protection against physical stress to the cells living on the scaffold surface. A dynamic cell seeding was developed and refined using the 3D culture system, which increased the 32% seeding efficiency of MC3T3 cells compared to the traditional static cell seeding method. The cell proliferation analysis demonstrated that HPSs could speed up cell growth in dynamic cell culture. The HPS with a honeycomb-like structure showed the highest inner pore velocity (CFD analysis) and achieved the fastest cell proliferation and the highest cell viability. Overall, our study, for the first time, developed a 3D printed HPS cell culture device with a uniformed porous structure, which can effectively facilitate cell adhesion and proliferation in the dynamic cultural environment, thereby could be considered an ideal carrier candidate.

Carbon Nanomaterials Modified Biomimetic Dental Implants for Diabetic Patients.

Dental implants are used broadly in dental clinics as the most natural-looking restoration option for replacing missing or highly diseased teeth. However, dental implant failure is a crucial issue for diabetic patients in need of dentition restoration, particularly when a lack of osseointegration and immunoregulatory incompetency occur during the healing phase, resulting in infection and fibrous encapsulation. Bio-inspired or biomimetic materials, which can mimic the characteristics of natural elements, are being investigated for use in the implant industry. This review discusses different biomimetic dental implants in terms of structural changes that enable antibacterial properties, drug delivery, immunomodulation, and osseointegration. We subsequently summarize the modification of dental implants for diabetes patients utilizing carbon nanomaterials, which have been recently found to improve the characteristics of biomimetic dental implants, including through antibacterial and anti-inflammatory capabilities, and by offering drug delivery properties that are essential for the success of dental implants.

Inhaled Edoxaban dry powder inhaler formulations: Development, characterization and their effects on the coagulopathy associated with COVID-19 infection.

Herein, we demonstrated the development and characterization of a dry powder inhaler (DPI) formulation of edoxaban (EDX); and investigated the in-vitro anticoagulation effect for the management of pulmonary or cerebral coagulopathy associated with COVID-19 infection. The formulations were prepared by mixing the inhalable micronized drug with a large carrier lactose and dispersibility enhancers, leucine, and magnesium stearate. The drug-excipient interaction was studied using X-Ray diffraction (XRD), Fourier transform infrared spectroscopy (FTIR), differential scanning calorimetry (DSC) and thermogravimetric analysis (TGA) methods. The drug and excipients showed no physical inter particulate interaction. The in-vitro drug aerosolization from the developed formulation was determined by a Twin Stage Impinger (TSI) at a flow rate of 60 ± 5 L /min. The amount of drug deposition was quantified by an established HPLC-UV method. The fine particle fraction (FPF) of EDX API from drug alone formulation was 7%, whereas the formulations with excipients increased dramatically to almost 7-folds up to 47%. The developed DPI formulation of EDX showed a promising in-vitro anticoagulation effect at a very low concentration. This novel DPI formulation of EDX could be a potential and effective inhalation therapy for managing pulmonary venous thromboembolism (VTE) associated with COVID-19 infection. Further studies are warranted to investigate the toxicity and clinical application of the inhaled EDX DPI formulation.

Endogenous nitric oxide-generating surfaces via polydopamine-copper coatings for preventing biofilm dispersal and promoting microbial killing.

INTRODUCTION: Peri-implantitis is a bacterially induced inflammatory disease which affects the hard and soft tissues around a dental implant. Microbial biofilm formation is an important causative factor in peri-implantitis. The aim of this study is to develop an effective multifunctional surface coating for antimicrobial property and to counteract oral biofilm-associated infections via a single polydopamine copper coating (PDAM@Cu) on titanium implant surface to regulate endogenous nitric oxide (NO) generation. METHODS: PDAM@Cu coatings were made with different concentrations of CuCl2 on titanium surfaces with a simple dip coating technique. Coatings were characterised to evaluate Cu concentrations as well as NO release rates from the coatings. Further, salivary biofilms were made on the coatings using Brain Heart Infusion (BHI) media in an anaerobic chamber. Biofilms were prepared with three different mixtures, one of which was saliva only, the second had an addition of sheep's blood, and the third was prepared with NO donors S-nitrosoglutathione (GSNO) and L-glutathione (GSH) in the mixture of saliva and blood to evaluate the effects of endogenously produced NO on biofilms. The effectiveness of coated surfaces on biofilms were assessed using four different methods, namely, crystal violet assay, scanning electron microscopy imaging, 2,3-bis (2-methoxy-4-nitro-5-sulfophenyl)-5-[(phenylamino) carbonyl]-2H-tetrazolium hydroxide (XTT) metabolic assay, and live/dead staining. RESULTS: NO release rates could be controlled with different Cu concentration in PDAM@Cu coatings. NO generated from the PDAM@Cu coatings effectively induced dispersal of biofilms shown by the reduction in biofilm biomass as well as reduced biofilm attachment in samples prepared with blood and NO donors. Cu ions released from the PDAM@Cu coatings resulted in killing of the dispersed bacteria, which was evidenced by the live/dead cell staining and reduced metabolic activity noted from the XTT assay. In contrast, samples prepared with saliva showed no significant reduction in biofilms, indicating the important effect of endogenously generated NO on biofilm dispersal. CONCLUSION: In conclusion, PDAM@Cu coatings with NO generating surfaces have a dual anti-biofilm function, with a synergistic effect on biofilm dispersal from regulated NO generation and bactericidal effects from Cu ions from the coatings.

The effects of TiO2 nanotube arrays with different diameters on macrophage/endothelial cell response and ex vivo hemocompatibility.

Percutaneous coronary intervention with stenting is the most widely adopted surgical technique for the treatment of coronary disease. However, in-stent restenosis (ISR) continues to cause serious concern. Improving the functionality of endothelial cells (ECs) is of importance for dealing with ISR. However, limited successes in in vivo studies imply that the underlying mechanism relating to the process of intervention with a material has not been well understood to date. Another determining factor that has long been underestimated in the design of stents is immune response dominated by macrophages (MΦs). In the present study, TiO2 nanotube arrays (TNAs) with different diameters were fabricated by anodization. The diameter could be precisely adjusted from 15 nm (Nano-15) to 120 nm (Nano-120). The effect of distinct nanodimensions on the behavior of ECs/MΦs and their crosstalk was investigated. The results indicated that Nano-15 not only promoted the initial vitality of ECs and function-related gene expression but also enhanced the activation and spread of MΦs. In addition, Nano-15 downregulated the gene expression of inflammatory cytokines and pro-inflammatory M1 markers, while upregulating the gene expression of pro-healing M2 markers, autophagy markers, and growth factors of MΦs. A further investigation conducted using a specialized specimen-conditioned medium (SCM) from MΦs/ECs disclosed that the SCM derived from Nano-15 was able to manipulate a favorable microenvironment to facilitate MΦ-EC crosstalk via downregulation of inflammation-related gene expression and upregulation of function-related gene expression in ECs. Moreover, we further investigated the hemocompatibility of specimens in an ex vivo animal model. The results indicated that both pure Ti and Nano-15 possess satisfactory hemocompatibility, as manifested by the few thrombi that formed around the specimens. Additionally, in comparison with Nano-120, Nano-15 can significantly alleviate platelet activation and manipulate a loose structure of fibrin fibers in a dynamic situation. This study suggests that the application of Nano-15 as a surface coating for coronary stents may be a promising strategy for preventing ISR.