Visualizing Extracellular Vesicles and Their Function in 3D Tumor Microenvironment Models

Extracellular vesicles (EVs) are cell-derived nanostructures that mediate intercellular communication by delivering complex signals in normal tissues and cancer. The cellular coordination required for tumor development and maintenance is mediated, in part, through EV transport of molecular cargo to resident and distant cells. Most studies on EV-mediated signaling have been performed in two-dimensional (2D) monolayer cell cultures, largely because of their simplicity and high-throughput screening capacity. Three-dimensional (3D) cell cultures can be used to study cell-to-cell and cell-to-matrix interactions, enabling the study of EV-mediated cellular communication. 3D cultures may best model the role of EVs in formation of the tumor microenvironment (TME) and cancer cell-stromal interactions that sustain tumor growth. In this review, we discuss EV biology in 3D culture correlates of the TME. This includes EV communication between cell types of the TME, differences in EV biogenesis and signaling associated with differing scaffold choices and in scaffold-free 3D cultures and cultivation of the premetastatic niche. An understanding of EV biogenesis and signaling within a 3D TME will improve culture correlates of oncogenesis, enable molecular control of the TME and aid development of drug delivery tools based on EV-mediated signaling.     

Strength retention of drawn self-reinforced polyglycolide rods and fixation properties of the distal femoral osteotomies with these rods. An experimental study on rats

Drawn self-reinforced polyglycolide (SR-PGA) rods, Ø 2 mm and 26 mm long, were implanted in the dorsal subcutaneus tissue of 16 rats. Osteotomies of the distal femur were fixed with SR-PGA rods (2 mm by 15 mm) in another 38 rats. The follow-up times varied from one week to one year. After sacrifice, three-point bending and shear tests were performed for subcutaneously placed rods. Radiological, histological, histomorphometrical, microradiographic, and oxytetracycline-fluorescence studies of osteotomized and intact control femora were also performed. At three weeks the flexural strength of the rods was 50% of the initial value, and the flexural modulus was 46% of the initial value. Five osteotomy specimens had to be excluded due to dislocation or non-union. One of the 33 evaluated osteotomy specimens showed signs of postoperative infection. Thirty-two osteotomies healed uneventfully. No gross signs of inflammatory or foreign-body reaction were observed. The amount of osteoid surface and active osteoid formation surface reached their highest value in the histomorphometrical analysis at 24 weeks. The present investigation demonstrated that the mechanical strength and fixation properties of the drawn SR-PGA rods are suitable for fixation of cancellous bone osteotomies in rats. The present article is the first report on the successful application of drawn SR-PGA rods for fixation of cancellous bone osteotomies.     

Repair of bone defects with absorbable membranes. A study on rabbits

Self-reinforced polyglycolic acid (SR-PGA) devices were developed in the mid-eighties, applied for fixation purposes and proved to be biocompatible. In this study SR-PGA membranes (10 x 10 mm) were used to augment defects on the medial aspect of distal femoral metaphysis in 31 New Zealand rabbits. Defects of 3.5 mm were either filled with autografts or left non-grafted. In a control group, no membranes were used. The rabbits were followed up for six, 12 and 24 weeks. Radiography, histology, oxytetracycline (OTC) fluorescence labelling and microradiography were used. Defects where membranes were used, healed by new bone formation. In some cases where polyglycolic acid (PGA) membranes were not used, defects were invaded by fibrous tissue. Membranes sometimes slipped away from their positions opposite to grafted defects. This study proved that the advantage of the use of PGA membranes could be taken in augmentation of cancellous bone defects in rabbits.     

Release of diclofenac sodium from polylactide-co-glycolide 80/20 rods

Due to inflammatory reactions complicating bioabsorbable devices, the aim of this study was to develop and characterize bioabsorbable implants with anti-inflammatory drug releasing properties. Polylactide-co- glycolide (PLGA) 80/20 was compounded with diclofenac sodium (DS) to produce rods. Thermal properties were analyzed using differential scanning calorimetry (DSC). Inherent viscosity (η_inh) was measured to evaluate the drug effect on the extrude polymer. Drug release measurements were performed using UV-spectrophotometer. Five parallel samples from each type of rods were examined, first at 6 hour intervals, then on daily basis, and later twice a week. DS was released in 110 days from thinner rods and in 150 days from thicker rods. Drug release comprised a starting peak, slow release phase, then a high release phase, and a burst release phase. DSC analysis showed that DS containing rods had crystallinity in their structure. In conclusions, it is feasible to combine PLGA 80/20 and DS by using melt extrusion. Released DS concentrations reached local therapeutic levels, but the release profile was complex and therapeutic levels were not reached all the time.     

Tissue reactions to bioabsorbable ciprofloxacin-releasing polylactide-polyglycolide 80/20 screws in rabbits’ cranial bone

The aim of this study was to assess tissue reactions to bioabsorbable self-reinforced ciprofloxacin-releasing polylactide/polyglycolide (SR-PLGA) 80/20 screws in rabbits' cranial bone. Two screws were implanted in each rabbit, one screw on either side of the sagittal suture (n = 28 rabbits). Animals were sacrificed after 2, 4, 8, 16, 24, 54 and 72 weeks, four animals per group. On histological examination the number of macrophages, giant cells, active osteoblasts and fibrous tissue layers were assessed and degradation of the screws was evaluated. At 2 weeks, the highest number of macrophages and giant cells were seen near the heads of the screws. After 4 and 8 weeks, the number of giant cells decreased but that of macrophages decreased from 16 weeks and on. Screws were surrounded by fibrous tissue capsule that progressively was growing in thickness by time. Active osteoblasts were seen around the shaft of the screws with the highest number seen at 4 weeks postoperatively. At 16 weeks, compact fragmentation of the screw heads was seen with macrophages seen inside the screw matrices. After 24 weeks, no polarization of the screws was seen. After one year, PLGA screws had been replaced by adipose tissue, fibrous tissue and "foamy macrophages" which had PLGA particles inside them. After 1(1/2) years, the amount of biomaterial remaining had decreased remarkably. The particles of biomaterial were inside "foamy macrophages." Ciprofloxacin-releasing SR-PLGA 80/20 screws elicited a mild inflammatory reaction but did not interfere with osteoblast activity. No complications were seen when implanted in cranial bone of rabbit.     

Poly(o-anisidine) on brass: Synthesis and corrosion behavior

The electrochemical synthesis of poly(o-anisidine) (POA) was achieved on brass (CuZn) electrode by applying two scan rates (50 and 20 mVs⁻¹). The synthesized polymer films were strongly adherent and homogeneous in both cases. Their corrosion performance was investigated by AC impedance spectroscopy (EIS) technique, anodic polarization plots and open circuit potential-time curves, in 3.5% NaCl solution. It was clearly seen that poly(o-anisidine) films provided a significant physical protection for longer exposure time. It was shown that polymer film coated at high scan rate (CuZn/POA-H) exhibited better barrier property against the attack of corrosive agents when compared with polymer film obtained at low scan rate (CuZn/POA-L). It was found out that poly(o-anisidine) film synthesized at high scan rate caused a significant increase in corrosion resistance by its catalytic behavior on formation of protective oxide layers on the surface in longer time.     

Synthetic Biology and Tissue Engineering: Toward Fabrication of Complex and Smart Cellular Constructs

Tissue engineering approaches, with the goals of replacing or recovering damaged or diseased tissues, or of reconstituting tissues in vitro for disease modeling and drug development, have the potential to make significant contributions to medicine. Advances in stem cell biology, biomaterial synthesis and characterization, and microscale technologies have made engineered tissues a reality. However, the classic tools used to build tissues in the lab do not allow for complete control of cell behaviors. More recently, synthetic biology principles have developed robust and versatile approaches to program cells with artificial genetic circuits, where cell behavior and function can be manipulated. At the interface between synthetic biology and tissue engineering, there is space for a new area of investigation where material engineering and cellular engineering complement and sustain each other. In this progress report, synthetic biology principles and how they have been used to engineer cells with potential to dictate cell behavior and function in tissue constructs of the future are briefly described. It is believed that this research area still needs further exploration to fully exploit synthetic biology to make smart and functional cellular constructs for therapeutic and in vitro applications.     

Biodegradable Implantable Sensors: Materials Design,Fabrication, and Applications

The ability to monitor diseases, therapies, and their effects on the body is a critical component of modern care and personalized medicine. Real time monitoring can be achieved by analyzing body fluids or by applying sensors on, or alternatively, inside the body. Implantable sensors, however, must be removed. Second removal procedures lead to further tissue damage, which can be a problem in tissues such as those of the central nervous system. The use of biodegradable sensors alleviates these problems since they do not require removal procedures. Recent advances in material science made it possible for all sensor components to be biodegradable. Small size and power of implants, and the limited selection of materials are the main constraints determining the capabilities of the biodegradable device. Thus, the design will be always a challenge exploring a trade-off among these parameters. Despite of the encouraging results illustrating that biodegradable sensors can be as accurate and reliable as commercially available nondegradable ones, biodegradable implantable sensors are still in their infancy. Significant advances made in this area are critically reviewed in this paper, and future prospects are highlighted.