In vivo acceleration of skin growth using a servo-controlled stretching device.

Tension is a principal force experienced by skin and serves a critical role in growth and development. Optimal tension application regimens may be an important component for skin tissue engineering and dermatogenesis. In this study, we designed and tested a novel servo-controlled skin-stretching device to apply predetermined tension and waveforms in mice. The effects of static and cyclical stretching forces were compared in 48 mice by measuring epidermal proliferation, angiogenesis, cutaneous perfusion, and principal growth factors using immunohistochemistry, real-time reverse transcriptase-polymerase chain reaction, and hyperspectral imaging. All stretched samples had upregulated epidermal proliferation and angiogenesis. Real-time reverse transcriptase-polymerase chain reaction of epidermal growth factor, transforming growth factor beta1, and nerve growth factor demonstrated greater expression in cyclically stretched skin when compared to static stretch. Hypoxia-induced factor 1alpha was significantly upregulated in cyclically stretched skin, but poststretch analysis demonstrated well-oxygenated tissue, collectively suggesting the presence of transient hypoxia. Waveform-specific mechanical loads may accelerate tissue growth by mechanotransduction and as a result of repeated cycles of temporary hypoxia. Further analysis of mechanotransduction signaling pathways may provide additional insight to improve skin tissue engineering methods and optimize our device.

Analysis of neuropeptides in stretched skin.

BACKGROUND: Mechanical forces modulate wound healing and scar formation through mechanotransduction. In response to mechanical stimulation, neuropeptides are released from peripheral terminals of primary afferent sensory neurons, influencing skin and immune cell functions and increasing vascular permeability, causing neurogenic inflammation. METHODS: A computer-controlled device was used to stretch murine skin. C57Bl6 mice (n = 26) were assigned to a cyclical square-wave tensile stimulation for 4 hours or continuous stimulation for 4 hours. Stretched skin was analyzed for expression of the neuropeptides, substance P and calcitonin gene-related peptide, their receptors (NK1R and calcitonin gene-related peptide receptor component protein), and growth factors (nerve growth factor, transforming growth factor beta1, vascular endothelial growth factor, and epidermal growth factor) using immunohistochemistry and real-time reverse-transcriptase polymerase chain reaction. RESULTS: Cyclical stimulation resulted in a significant increase in expression of neuropeptides and growth factors, whereas the corresponding peptide receptors were down-regulated. Transcription of neuropeptide mRNA was elevated in stretched skin, which proves that neuropeptides are released from not only peripheral terminals of nerve fibers but also resident skin cells. CONCLUSIONS: The authors' results suggest that skin stretching may alter cell physiology by stimulating neuropeptide expression, and that cyclical mechanical force may be more effectively stimulating mechanosensitive nociceptors or mechanoreceptors (mechanosensors) on cells.

The mechanism of action of the vacuum-assisted closure device.

BACKGROUND: The vacuum-assisted closure device is widely used clinically, yet its mechanisms of action are incompletely understood. In this study, the authors designed a partially splinted full-thickness murine vacuum-assisted closure model to better understand the mechanism of action of the vacuum-assisted closure device. METHODS: Full-thickness wounds (n = 10 per group) were excised in diabetic mice and treated with the vacuum-assisted closure device or its isolated components: an occlusive dressing, subatmospheric pressure at 125 mmHg (suction), and a polyurethane foam without and with downward compression. Results were quantified with a two-dimensional immunohistochemical staging system based on blood vessel density (CD31) and cell proliferation (Ki67) 7 days after wounding. Microscopic strain was measured by fixing in situ all dressing modalities. RESULTS: Wounds exposed to polyurethane foam in compressed and uncompressed dressings or to the vacuum-assisted closure device showed a 2-fold increase in vascularity compared with the occlusive dressing group (p < 0.05). The vacuum-assisted closure device in addition stimulated cell proliferation, with up to 82 percent Ki67-positive nuclei, compared with the other groups. Direct measurements of wound surface deformations showed significant microstrains in the vacuum-assisted closure and foam in compressed dressing groups (60 percent and 16 percent, respectively) compared with all other groups. CONCLUSIONS: These data provide profound insights into the mechanism of action of the vacuum-assisted closure device, providing an explanation for the increases in wound bed vascularity and cell proliferation based on its components. Results suggest that the vascular response is related to the polyurethane foam, whereas tissue strains induced by the vacuum-assisted closure device stimulated cell proliferation.

Tensile forces stimulate vascular remodeling and epidermal cell proliferation in living skin.

OBJECTIVES: To quantify tissue remodeling induced by static and cyclical application of tensional forces in a living perfused tissue. BACKGROUND: Cells are able to respond to mechanical cues from the environment and can switch between proliferation and quiescence. However, the effects of different regimens of tension on living, perfused skin have not been characterized. METHODS: The ears of living rats were mechanically loaded by applying tensile forces (0.5 Newtons) either statically or cyclically and then analyzing tissue responses using in vivo microscopy, immunohistochemistry, and corrosion casting. RESULTS: Quantitative immunohistochemistry showed that in the static group (4-day continuous tension) there was up to 4-fold increase in cellular proliferation in the epidermis after 4 days and a 2.8-fold increase in the vascularity in the dermis that peaked after 2 days. Comparable effects could be achieved in just 8 hours using a cyclic loading protocol. We also modeled the resultant stress produced in the ear using a linear finite element model and demonstrated a correlation between the level of applied stress and both epidermal cell proliferation and blood vessel density. CONCLUSIONS: Mechanical forces stimulate cell proliferation and vascular remodeling in living skin. As cell growth and vascular supply are critical to wound healing and tissue expansion, devices applying controlled mechanical loads to tissues may be a powerful therapy to treat tissue defects.