al Low-Intensity Vibration and Wound Healing Wound healing assays Wound healing was assessed histologically using our previously published assays of re-epithelialization, granulation tissue formation, collagen deposition, and angiogenesis using hematoxylin and eosin, PubMed ID:http://www.ncbi.nlm.nih.gov/pubmed/19632594 Masson’s Trichrome and CD31 stained cryosections. For all wound healing analyses, digital images were obtained using a Nikon Instruments Eclipse 80i microscope with a 206/0.75 objective, a DS-Fi1 digital camera, and NIS Elements software. Tissue preparation. Two wounds per mouse were collected and sectioned from one edge to well past the center. Sections were then selected from the center of the wound by microscopic assessment. Three 10-mm sections judged to be at the actual center of the wound were used for re-epithelialization and granulation tissue thickness Sodium laureth sulfate measurements. Adjacent three 10-mm sections were used for trichrome staining, CD31 staining, and inflammatory cell staining. Re-epithelialization and granulation tissue thickness. Wound re-epithelialization was measured by mor- hypothesis of this study was that LIV improves the delayed wound healing in diabetic mice by promoting a pro-angiogenic and prohealing wound environment. Methods Animals Diabetic db/db mice, which are widely used as a model of delayed healing, were obtained from Jackson Laboratories. Experiments were performed on male mice 1216 week-old. Only mice with fasting blood glucose.250 mg/dl were used. All procedures involving animals were approved by the Animal Care Committee at the University of Illinois at Chicago. All animals were housed under standard conditions and treated according to the Guide for the Care and Use of Laboratory Animals of the NIH. Excisional wounding Mice were subjected to excisional wounding as described previously. Briefly, mice were anesthetized with isoflurane and their dorsum was shaved and cleaned with alcohol. Four 8 mm wounds were made on the back of each mouse with a dermal biopsy punch and covered with Tegaderm to keep the wounds moist and maintain consistency with treatment of human wounds. Wounds were harvested at 7 and 15 days post-wounding. Low-intensity vibration Following wounding, mice were randomly assigned to the LIV or a non-vibration sham treatment. For the LIV, mice were placed in an empty cage directly on the vibrating plate, and LIV was applied vertically at 45 Hz with peak acceleration of 0.4 g for 30 min per day for 5 days/week starting on the day of wounding. The non-vibrated sham controls were similarly placed in a separate empty cage but were not subjected to LIV. The mechanical signals were calibrated using an accelerometer attached to the inside surface of the bottom of the cage, so that the signals produced were indeed those transmitted to the feet of the mice. In addition, the amplitude of the vibrations are small enough that the cage does not move relative to the plate and the vibrations of the plate and the cage are in sync. phometric analysis PubMed ID:http://www.ncbi.nlm.nih.gov/pubmed/19630872 of wound sections. Sections 
taken from the center of the wound were stained with H&E. The distance between the wound edges, defined by the distance between the first hair follicle encountered at each end of the wound, and the distance that the epithelium had traversed into the wound, were measured using image analysis software. The percentage of reepithelialization and granulation tissue thickness was calculated for three sections per wound and was averaged over sections to provide a representative valu
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