A preliminary In Vitro viability study of an electrically active hernia mesh on mouse fibroblasts.

Hernias occur when part of an organ, typically the intestines, protrudes through a disruption of the fascia in the abdominal wall, leading to patient pain, discomfort, and surgical intervention. Over one million hernia repair surgeries occur annually in the USA, but globally, hernia surgeries can exceed 20 million. Standard practice includes hernia repair mesh to help hold the compromised tissue together, depending on where the fascial disruption is located and the patient's condition. However, the recurrence rate for hernias after using the most common type of hernia mesh, synthetic, is currently high. Physiological-level electrical stimulation (ES) has shown beneficial effects in improving healing in soft tissue regeneration. Piezoelectric materials can produce low-level electrical signals from mechanical loading to help speed healing. Combining the novelty of piezo elements to create an electrically active hernia repair mesh for faster healing prospects is explored in this study through simulated transcutaneous mechanical loading of the piezo element with therapeutic ultrasound. A tissue phantom was developed using Gelatin #0 and Metamucil® to better simulate a clinical application of the therapeutic ultrasound loading modality. The cellular viability of varying ultrasound intensities and temporal effects was analyzed. Overall, minimal cytotoxicity was observed across all experimental groups during the ultrasound intensity and temporal viability studies.

Integration of clinical perspective into biomimetic bioreactor design for orthopedics.

The challenges to accommodate multiple tissue formation metrics in conventional bioreactors have resulted in an increased interest to explore novel bioreactor designs. Bioreactors allow researchers to isolate variables in controlled environments to quantify cell response. While current bioreactor designs can effectively provide either mechanical, electrical, or chemical stimuli to the controlled environment, these systems lack the ability to combine all these stimuli simultaneously to better recapitulate the physiological environment. Introducing a dynamic and systematic combination of biomimetic stimuli bioreactor systems could tremendously enhance its clinical relevance in research. Thus, cues from different tissue responses should be studied collectively and included in the design of a biomimetic bioreactor platform. This review begins by providing a summary on the progression of bioreactors from simple to complex designs, focusing on the major advances in bioreactor technology and the approaches employed to better simulate in vivo conditions. The current state of bioreactors in terms of their clinical relevance is also analyzed. Finally, this review provides a comprehensive overview of individual biophysical stimuli and their role in establishing a biomimetic microenvironment for tissue engineering. To date, the most advanced bioreactor designs only incorporate one or two stimuli. Thus, the cell response measured is likely unrelated to the actual clinical performance. Integrating clinically relevant stimuli in bioreactor designs to study cell response can further advance the understanding of physical phenomenon naturally occurring in the body. In the future, the clinically informed biomimetic bioreactor could yield more efficiently translatable results for improved patient care.

Reverse engineering of fatty acid-tolerant Escherichia coli identifies design strategies for robust microbial cell factories.

Adaptive laboratory evolution is often used to improve the performance of microbial cell factories. Reverse engineering of evolved strains enables learning and subsequent incorporation of novel design strategies via the design-build-test-learn cycle. Here, we reverse engineer a strain of Escherichia coli previously evolved for increased tolerance of octanoic acid (C8), an attractive biorenewable chemical, resulting in increased C8 production, increased butanol tolerance, and altered membrane properties. Here, evolution was determined to have occurred first through the restoration of WaaG activity, involved in the production of lipopolysaccharides, then an amino acid change in RpoC, a subunit of RNA polymerase, and finally mutation of the BasS-BasR two component system. All three mutations were required in order to reproduce the increased growth rate in the presence of 20 mM C8 and increased cell surface hydrophobicity; the WaaG and RpoC mutations both contributed to increased C8 titers, with the RpoC mutation appearing to be the major driver of this effect. Each of these mutations contributed to changes in the cell membrane. Increased membrane integrity and rigidity and decreased abundance of extracellular polymeric substances can be attributed to the restoration of WaaG. The increase in average lipid tail length can be attributed to the RpoCH419P mutation, which also confers tolerance to other industrially-relevant inhibitors, such as furfural, vanillin and n-butanol. The RpoCH419P mutation may impact binding or function of the stringent response alarmone ppGpp to RpoC site 1. Each of these mutations provides novel strategies for engineering microbial robustness, particularly at the level of the microbial cell membrane.