Platelet Rich Fibrin (PRF) is a biological and innovative therapeutic which has important role in regenerative injured tissue. The aims of this study were to prepare lyophilized-PRF and determine its applications during wound healing, depending on histopathological and immunohistochemistry parameters. Six healthy, adult local breed dogs were used, the animals were randomly divided into 2 experimental main groups (treated and control) 6 animals for each group, ten ml of whole blood was aspirated from the same animal for platelet rich fibrin preparation and centrifugation of this fresh blood at 3000 rpm for 10 minutes was performed, the resulted PRF layer was removed to prepare lyophilized PRF by freezing and was stored at −80C∘ then the sample was freeze-dried for 12 hours by using  Labconco lyophilizer at −51C∘ Surgical full thickness incision wound of 5cm length was performed under general anesthesia on back of dogs under aseptic technique, treated group Lyophilized A-PRF (Right side) was applied subcutaneously while normal saline 0.9% was applied instead of Lyophilized A-PRF in the control group (Left side). The wound was closed using surgical silk suture.  The histological criteria for wound healing, inflammatory cell infiltration, re-epithelialization, and granulation tissue formation were evaluated according to the scoring system (0-4), Furthermore, the results of quantitative analysis of immunohistochemistry of PDGF expression showed that the group treated with Lyph-PRF was moderate, intense and mild cytoplasmic staining reactivity on the first, third and seventh days respectively after PO compared to the control.