RESUMEN
The advent of recombinant DNA technology fundamentally altered the drug discovery landscape, replacing traditional small-molecule drugs with protein and peptide-based biologics. Being susceptible to degradation via the oral route, biologics require comparatively invasive injections, most commonly by intravenous infusion (IV). Significant academic and industrial efforts are underway to replace IV transport with subcutaneous delivery by wearable infusion devices. To further complement the ease-of-use and safety of disposable infusion devices, surface disinfection of the drug container can be automated. For ease of use, the desired injector is a combination device, where the drug is inside the injector as a single solution combination device. The main obstacle of the desired solution is the inability to sterilize both injector and drug in the same chamber or using the same method (Gamma for the drug and ETO for the injector). This leads to the assembly of both drug container and injector after sterilization, resulting in at least one transition area that is not sterilized. To automate the delivery of the drug to the patient, a disinfection step before the drug delivery through the injector is required on the none-sterilized interface. As an innovative solution, the autoinjector presented here is designed with a single ultraviolet light-emitting diode (UV LED) for surface disinfection of the drug container and injector interface. In order to validate microbial disinfection similar to ethanol swabbing on the injector, a bacterial 3 or 6 log reduction needed to be demonstrated. However, the small disinfection chamber surfaces within the device are incapable of holding an initial bacterial load for demonstrating the 3 or 6 log reduction, complicating the validation method, and presenting a dilemma as to how to achieve the log reduction while producing real chamber conditions. The suggested solution in this paper is to establish a correlation model between the UV irradiance distribution within the disinfection chamber and a larger external test setup, which can hold the required bacterial load and represents a worse-case test scenario. Bacterial log reduction was subsequently performed on nine different microorganisms of low to high UV-tolerance. The procedure defined herein can be adopted for other surface or chamber disinfection studies in which the inoculation space is limited.
RESUMEN
There is a need to further explore the convergence of mechanobiology and dimensionality with systematic investigations of cellular response to matrix mechanics in 2D and 3D cultures. Here, a semisynthetic hydrogel capable of supporting both 2D and 3D cell culture is applied to investigate cell response to matrix modulus and ligand density. The culture materials are fabricated from adducts of polyethylene glycol (PEG) or PluronicF127 and fibrinogen fragments, formed into hydrogels by free-radical polymerization, and characterized by shear rheology. Control over the modulus of the materials is accomplished by changing the concentration of synthetic PEG-diacrylate crosslinker (0.5% w/v), and by altering the molecular length of the PEG (10 and 20 kDa). Control over ligand density is accomplished by changing fibrinogen concentrations from 3 to 12 mg mL-1 . In 2D culture, cell motility parameters, including cell speed and persistence time are significantly increased with increasing modulus. In both 2D and 3D culture, cells express vinculin and there is evidence of focal adhesion formation in the high stiffness materials. The modulus- and ligand-dependent morphogenesis response from the cells in 2D culture is contradictory to the same measured response in 3D culture. In 2D culture, anchorage-dependent cells become more elongated and significantly increase their size with increasing ligand density and matrix modulus. In 3D culture, the same anchorage-dependent cells become less spindled and significantly reduce their size in response to increasing ligand density and matrix modulus. These differences arise from dimensionality constraints, most notably the encapsulation of cells in a non-porous hydrogel matrix. These insights underscore the importance of mechanical properties in regulating cell morphogenesis in a 3D culture milieu. The versatility of the hydrogel culture environment further highlights the significance of a modular approach when developing materials that aim to optimize the cell culture environment.