Outcome measures
TNF-α, necrotic
STERILE, NECROTIC
Subjects were asked to come twice a week, for 4 consecutive weeks. Every subject was given standard treatment for DFU, namely washing the wound with normal saline (0.9% NaCl) and removing necrotic tissue using sharp debridement method.After wound debridement, measurement of wound area was conducted using planimetric method and digital photography with two rulers. The entire surface of the wound and rulers were photographed using a Canon PowerShot G7X digital camera (20.20 megapixels). The image was taken perpendicularly to form an angle of 90° to the wound surface at a distance of 30 cm, with good room lighting. The photos were then transferred to a computer. Photos were processed using ImageJ. At first, the wound size was calibrated based on the ruler scale on the edge of the wound. Then the edges of the wound were traced using mouse cursor for the calculation of the surface area. In addition to the wound edges, the amount of granulation tissue was also traced using mouse and the area was calculated. This was repeated several times at different times to see the progress of wound healing. The granulation index was calculated by dividing the granulation tissue area by the measured wound area.Subsequently, the wound swab was taken to examine the number and pattern of aerobic bacteria colonization. Wound swabs were taken with Puritan Calcium Alginate Swab 25-806 1PA (Puritan, USA) using the Levine wound swab technique and a sterile square marker made of metal. The swab tip was immediately inserted into a tube containing 5 mL of phosphate-buffered saline (PBS) and immediately delivered to the Eijkman Microbiology Laboratory.Wound fluid collection for the examination of LL-37, IL-1α, and TNF-α was performed using Whatman