miR-200b & miR-21 in Diabetic Wounds

The study is designed as a 14 week prospective observational study where patients with
diabetic wounds visiting the Ohio State University Comprehensive Wound Center (CWC) will be
enrolled. Patients enrolled will be followed for 14 weeks including 4 study visits, initial
visit (week 0), study visit 2 (week 2), study visit 3 (week 4) and will return for a final
follow-up study visit 4 (week 14). Standardized Care Outline will be per physician
discretion. The initial visit will take place during the patients regularly scheduled CWC
visit and the following will take place: Informed consent will be obtained, Baseline
demographics, medical history, current labs and medications will be recorded, Wound data will
be recorded, Ankle brachial index will be completed for patients with leg wounds if not
already completed per Standard of Care (SoC) since onset of the wound, Wound photographs for
digital planimetry to measure wound area, Wound fluid will be collected (mir200b arm only):
Fluid will be collected from filter paper method or aspiration method. Wound fluid collection
will take place before debridement and biopsies.The NPWT sponge will be collected (mir200b
and miR-21 arm): Wound macrophages, immune cells (miR-21) and endothelial cells (miR-200b)
will be isolated from wound cells obtained from the sponge dressing of the NWPT. The NWPT
delivers negative pressure to help promote wound healing. NWPT is a routinely applied to
chronic wounds as therapeutic modality. The foam dressing to be collected is discarded as
biological waste at the regular CWC visits. Two 3 mm punch tissue biopsies will be obtained
from the patient's physician. One biopsy will be used for LCM (wound-edge endothelial cell
capture) and immunohistochemistry studies and the other biopsy will be used to determine if
the wound is infected using quantitative bacterial cultures and scanning electron microscopy
for biofilm structure detection.

Patient will return for study visit 2 where the following will happen: Patient data will be
collected including: medication review, adverse event review wound data will be recorded,
Wound photographs for digital planimetry to measure wound area, Wound fluid (miR-200b arm
only) will be collected, NPWT will be collected (miR-200b and miR-21 arms) and two 3 mm punch
tissue biopsies will be obtained from the patient's physician. Note- These biopsies will be
optional. At the beginning of enrollment the patient will be asked if they would like these
biopsies to be obtained. The research personnel will make note of the patients' decision. If
the wound shows signs of healing and does not show signs of infection (i.e. the wound has not
stalled in healing for 2 or more weeks or become larger in size) or the wound has been
declared healed by their physician, the biopsies will not be obtained. Patients will return
for study visit 3. The following will take place: Patient data will be collected including:
medication review, adverse event review, wound measurements, etiology, location and duration
will be recorded, Wound photographs for digital planimetry to measure wound area, Wound fluid
will be collected (miR-200b arm only), NPWT will be collected (miR-200b and miR-21 arms), Two
3 mm punch tissue biopsies will be obtained from the patient's physician. Note- if the
subjects wound has been declared healed by their physician, the biopsies will not be
obtained. Patients will return for study visit 4 where the following will take place: Medical
Chart review to determine final status of the wound as healed, healing or not healed, Wound
data will be recorded, Adverse Event review, Wound photographs for digital planimetry to
measure wound area. The risks associated with this research study are low. There are no
therapeutic interventions or medications as part of this study to be administered depending
on the amount of sensation the subject has at the site of the wound. The wound site will be
numbed by applying local anesthesia as appropriate to the area being biopsied. There may be
some discomfort with the tissue biopsy procedure. The wound site will be numbed by applying
local anesthesia as appropriate to the area being biopsied. The amount of pain that occurs
with wound biopsy will vary from person to person, but all reasonable efforts will be made to
minimize pain. The biopsy procedure is done within the boundaries of the existing wound to
avoid giving the patient a separate new wound. Bleeding is a possible complication, but the
risk is low for the small biopsies and is reduced by using local anesthetics with
epinephrine, and silver nitrate sticks are available in each room to cauterize biopsy sites
as needed. Infection is also a potential risk; however, wound tissue biopsies are routinely
performed as the standard of care in the CWC to diagnose wound infection. The biopsy site
will be appropriately monitored for infection by the physician managing the wound during
routine wound clinic visits. The ABI test may result in temporary discomfort around the ankle
or foot when the cuff is inflated, but does not present any further physical or medical
risks. The wound fluid collection is non-invasive and does not present any risk. Study
participants will not benefit directly from participation in this study. This research
information has the potential of providing considerable benefits to wound care by identifying
a key player that influences the closure of chronic diabetic wounds. Such knowledge should
help develop novel biomarker and or miR-directed therapeutic strategies.

Inclusion Criteria:

- Ages 18 to 80 years old

- Willing and able to provide informed consent

- Willing and able to comply with protocol instructions, including all biopsies and
study visits

- Type 2 Diabetes Mellitus

- Chronic wounds (open >30d)

- Diabetic foot ulcers, Wagner 1 or 2

- Target wound is non-infected (infection status will be determined by a culture report)

- The subject must not have been on antibiotics for the target wound within 2 weeks
prior to enrollment

- Subjects with Negative Pressure Wound Therapy (NPWT or also called a wound vac)

- Note: Applies only to the miR-21 arm

Exclusion Criteria:

- Revascularization surgery on the target wound within 60 days prior to enrollment

- Inadequate arterial supply, as evidenced by any of the following:

- Transcutaneous Oxygen Measurement (TcOM) < 30mmg

- Ankle Brachial Index (ABI) <0.7 or 1.3

- Toe Brachial Index (TBI) <0.6

- Subjects with marked immunodeficiency (HIV/AIDS, organ transplant patients and
actively being treated for cancer)

- Trauma wounds

- Wounds closed or to be closed by flap or graft coverage

- Women who are pregnant