Collagen Nanoyarns: Hierarchical Three-Dimensional Biomaterial Constructs.

Hierarchical fibrous scaffolds (HFS) consist of nanoscale fibers arranged in larger macroscale structures, much in the same pattern as in native tissue such as tendon and bone. Creation of continuous macroscale nanofiber yarns has been made possible using modified electrospinning set-ups that combine electrospinning with techniques such as twisting, drawing, and winding. In this paper, a modified electrospinning setup was used to create continuous yarns of twisted type I collagen nanofibers, also known as collagen nanoyarns (CNY), from collagen solution prepared in acetic acid. Fabricated CNYs were cross-linked and characterized using SEM imaging and mechanical testing, while denaturation of collagen and dissolution of the scaffolds were assessed using circular dichroism (CD) and UV-vis spectroscopy, respectively. HeLa cells were then cultured on the nanoyarns for 24 h to assess cell adhesion on the scaffolds. Scanning electron micrographs revealed a twisted nanofiber morphology with an average nanofiber diameter of 213 ± 60 nm and a yarn diameter of 372 ± 23 µm that shrank by 35% after covalent cross-linking. Structural denaturation assessment of native collagen using circular dichroism (CD) spectroscopy showed that 60% of the triple-helical collagen content in CNYs was retained. Cross-linking of CNYs significantly improved their mechanical properties as well as stability in buffered saline with no sign of degradation for 14 days. In addition, CNY strength and stiffness increased significantly with cross-linking although in the wet state, significant loss in these properties, with a corresponding increase in elasticity, was observed. HeLa cells cultured on cross-linked CNYs for 24 h adhered to the yarn surface and oriented along the nanofiber alignment axis, displaying the characteristic spindle-like morphology of cells grown on surfaces with aligned topography. Collectively, the results demonstrate the promising potential of collagen nanoyarns as a new class of shapable biomaterial scaffold and building block for generating macroscale fiber-based tissues.

Low-frequency (<100 kHz), low-intensity (<100 mW/cm(2)) ultrasound to treat venous ulcers: a human study and in vitro experiments.

The purpose of this study was to examine whether low frequency (<100 kHz), low intensity (<100 mW/cm(2), spatial peak temporal peak) ultrasound can be an effective treatment of venous stasis ulcers, which affect 500 000 patients annually costing over $1 billion per year. Twenty subjects were treated with either 20 or 100 kHz ultrasound for between 15 and 45 min per session for a maximum of four treatments. Healing was monitored by changes in wound area. Additionally, two in vitro studies were conducted using fibroblasts exposed to 20 kHz ultrasound to confirm the ultrasound's effects on proliferation and cellular metabolism. Subjects receiving 20 kHz ultrasound for 15 min showed statistically faster (p < 0.03) rate of wound closure. All five of these subjects fully healed by the fourth treatment session. The in vitro results indicated that 20 kHz ultrasound at 100 mW/cm(2) caused an average of 32% increased metabolism (p < 0.05) and 40% increased cell proliferation (p < 0.01) after 24 h when compared to the control, non-treated cells. Although statistically limited, this work supports the notion that low-intensity, low-frequency ultrasound is beneficial for treating venous ulcers.

Manipulating multifaceted microbubble shell composition to target both TRAIL-sensitive and resistant cells.

This study represents the first attempt to combine surface TRAIL expression and doxorubicin co-encapsulation in a single drug delivery agent in the form of ultrasound-responsive microbubbles that shatter into fragments, or nanoshards, in an ultrasound beam. We compare customized microbubbles of different polymeric shell compositions, and investigate the effect of both shell composition and incorporation of doxorubicin on action against TRAIL-sensitive MDA-MB-231 and TRAIL-resistant MCF7 human breast adenocarcinoma cells. Ligation of TRAIL only significantly impacted MDA-MB-231 cells predominantly by apoptosis, and had minimal effect on MCF12A (normal control) cells. For all shell types, nanoshards had a greater effect (apoptotic death ranging from approximately 25% for 1 wt % LipidPEG to 50% for 100% PLA), reflecting the greater surface area and larger number of particles that ultrasound generates. Encapsulation of doxorubicin generated necrosis in all cell lines, but PEGylation produced less effective necrosis in all cell lines. Co-encapsulation of doxorubicin within the contrast agent shell increased MDA-MB-231 cell death to approximately 40-80%, representing a marked increase over TRAIL alone, reflecting the dramatic effect of shell composition. Additionally, shells that co-encapsulated TRAIL and doxorubicin resulted in approximately 30-60% death in TRAIL-resistant MCF7 human breast adenocarcinoma cells, compared with little apoptotic response in these cells from shells encapsulating TRAIL alone, demonstrating the sensitization effect of the drug. This work has resulted in production of a library of effective ultrasound-triggered, minimally immunogenic, targeted drug delivery agents for potential use in cancer therapy, and represents a promising multifaceted treatment to better serve the population with solid tumors. © 2018 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 106A: 1903-1915, 2018.