Vitrification, a kinetic process of liquid solidification into glass, poses many potential benefits for tissue cryopreservation including indefinite storage, banking, and facilitation of tissue matching for transplantation. To date, however, successful rewarming of tissues vitrified in VS55, a cryoprotectant solution, can only be achieved by convective warming of small volumes on the order of one mL. Successful rewarming requires both uniform and fast rates in order to reduce thermal mechanical stress and cracks, and to prevent rewarming phase crystallization. Here we present a scalable nanowarming technology for 1 to 80 mL samples using radiofrequency-excited mesoporous silica coated iron oxide nanoparticles in VS55. Advanced imaging including Sweep Imaging with Fourier Transform and micro computed tomography were used to verify loading and unloading of VS55 and nanoparticles and successful vitrification of human dermal fibroblast cells, porcine arteries, and porcine aortic heart valve leaflet tissue. Nanowarming was then used to demonstrate uniform and rapid rewarming at > 130 °C/min in both physical (1 to 80 mL) and biological systems (1 to 50 mL). Nanowarming yielded viability that matched control and/or exceeded gold standard convective warming in 1 to 50 mL systems, and improved viability compared to slow warmed (crystallized) samples. Finally, biomechanical testing displayed no significant biomechanical property changes in blood vessel length or elastic modulus after nanowarming compared to untreated fresh control porcine arteries. In aggregate, these results demonstrate new physical and biological evidence that nanowarming can improve the outcome of vitrified cryogenic storage of tissues in larger sample volumes.

Aims Most studies of biofilm effects on dental materials use single-species biofilms, or consortia. Microcosm biofilms grown directly from saliva or plaque are much more diverse, but difficult to characterize. We used the Human Oral Microbial Identification Microarray (HOMIM) to validate a reproducible oral microcosm model. Methods and Results Saliva and dental plaque were collected from adults and children. Hydroxyapatite and dental composite disks were inoculated with either saliva or plaque, and microcosm biofilms were grown in a CDC biofilm reactor. In later experiments, the reactor was pulsed with sucrose. DNA from inoculums and microcosms were analyzed by HOMIM for 272 species. Microcosms included about 60% of species from the original inoculum. Biofilms grown on hydroxyapatite and composites were extremely similar. Sucrose-pulsing decreased diversity and pH, but increased the abundance of Streptococcus and Veilonella. Biofilms from the same donor, grown at different times, clustered together. Conclusions This model produced reproducible microcosm biofilms that were representative of the oral microbiota. Sucrose induced changes associated with dental caries. Significance and Impact of the Study This is the first use of HOMIM to validate an oral microcosm model that can be used to study the effects of complex biofilms on dental materials.

This report describes the design, fabrication, and testing of a cross-flow filtration microdevice, for the continuous extraction of blood plasma from a circulating whole blood sample in a clinically relevant environment to assist in continuous monitoring of a patient’s inflammatory response during cardiac surgeries involving cardiopulmonary bypass (CPB) procedures (about 400 000 adult and 20 000 pediatric patients in the United States per year). The microfiltration system consists of a two-compartment mass exchanger with two aligned sets of PDMS microchannels, separated by a porous polycarbonate (PCTE) membrane. Using this microdevice, blood plasma has been continuously separated from blood cells in a real-time manner with no evidence of bio-fouling or cell lysis. The technology is designed to continuously extract plasma containing diagnostic plasma proteins such as complements and cytokines using a significantly smaller blood volume as compared to traditional blood collection techniques. The microfiltration device has been tested using a simulated CPB circulation loop primed with donor human blood, in a manner identical to a clinical surgical setup, to collect plasma fractions in order to study the effects of CPB system components and circulation on immune activation during extracorporeal circulatory support. The microdevice, with 200 nm membrane pore size, was connected to a simulated CPB circuit, and was able to continuously extract ~15% pure plasma volume (100% cell-free) with high sampling frequencies which could be analyzed directly following collection with no need to further centrifuge or modify the fraction. Less than 2.5 ml total plasma volume was collected over a 4 h sampling period (less than one Vacutainer blood collection tube volume). The results tracked cytokine concentrations collected from both the reservoir and filtrate samples which were comparable to those from direct blood draws, indicating very high protein recovery of the microdevice. Additionally, the cytokine concentration increased significantly compared to baseline values over the circulation time for all cytokines analyzed. The high plasma protein recovery (over 80%), no indication of hemolysis and low level of biofouling on the membrane surface during the experimental period (over 4 h) were all indications of effective and reliable device performance for future clinical applications. The simple and robust design and operation of these devices allow operation over a wide range of experimental flow conditions and blood hematocrit levels to allow surgeons and clinicians autonomous usage in a clinical environment to better understand the mechanisms of injury resulting from cardiac surgery, and allow early interventions in patients with excessive postoperative complications to improve surgical outcomes. Ultimately, monolithic integration of this microfiltration device with a continuous microimmunoassay would create an integrated microanalysis system for tracking inflammation biomarkers concentrations in patients for point-of-care diagnos...

Dental caries, i.e., tooth decay mediated by bacterial activity, is the most widespread chronic disease worldwide. Carious lesions are commonly treated using dental resin composite restorations. However, resin composite restorations are prone to recurrent caries, i.e., reinfection of the surrounding dental hard tissues. Recurrent caries is mainly a consequence of waterborne and/or biofilm-mediated degradation of the tooth-restoration interface through hydrolytic, acidic and/or enzymatic challenges. Here we use amphipathic antimicrobial peptides to directly coat dentin to provide resin composite restorations with a 2-tier protective system, simultaneously exploiting the physicochemical and biological properties of these peptides. Our peptide coatings modulate dentin's hydrophobicity, impermeabilize it, and are active against multispecies biofilms derived from caries-active individuals. Therefore, the coatings hinder water penetration along the otherwise vulnerable dentin-restoration interface, even after in vitro aging, and increase its resistance against degradation by water, acids, and saliva. Moreover, they do not weaken the resin composite restorations mechanically. The peptide-coated highly-hydrophobic dentin is expected to notably improve service life of resin composite restorations and to enable the development of entirely hydrophobic restorative systems. The peptide coatings were also antimicrobial and thus, they provide a second tier of protection preventing re-infection of tissues in contact with restorations.

Objective To develop a method for quantifying leakage in composite resin restorations after curing, using non-destructive X-ray micro-computed tomography (micro-CT) and image segmentation. Methods Class-I cavity preparations were made in 20 human third molars, which were divided into 2 groups. Group I was restored with Z100 and Group II with Filtek LS. Micro-CT scans were taken for both groups before and after they were submerged in silver nitrate solution (AgNO3 50%) to reveal any interfacial gap and leakage at the tooth restoration interface. Image segmentation was carried out by first performing image correlation to align the before- and after-treatment images and then by image subtraction to isolate the silver nitrate penetrant for precise volume calculation. Two-tailed Student’s t-test was used to analyze the results, with the level of significance set at p<0.05. Results All samples from Group I showed silver nitrate penetration with a mean volume of 1.3 ± 0.7 mm3. In Group II, only 2 out of the 10 restorations displayed infiltration along the interface, giving a mean volume of 0.3 ± 0.3 mm3. The difference between the two groups was statistically significant (p < 0.05). The infiltration showed non-uniform patterns within the interface. Significance We have developed a method to quantify the volume of leakage using non-destructive micro-CT, silver nitrate infiltration and image segmentation. Our results confirmed that substantial leakage could occur in composite restorations that have imperfections in the adhesive layer or interfacial debonding through polymerization shrinkage. For the restorative systems investigated in this study, this occurred mostly at the interface between the adhesive system and the tooth structure.