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Background: Effectively managing acne scars while minimizing consequences is still a challenging task. The primary determinant in selecting and continuing therapy for a specific device is stimulating collagen production and dermal remodeling with the fewest possible complications. Objective: To evaluate the clinical and immunohistochemical results of ablative fractional carbon dioxide (FCO₂) laser versus fractional non-ablative diode laser for treating facial acne scars. Methods and Materials: Thirty patients with atrophic acne scars were included in a split-face comparative study. Right and left facial sides received three sessions of FCO₂ and diode laser, respectively, at one-month intervals. One month after the third session, patients were evaluated using photographs, the Goodman and Baron qualitative and quantitative global scarring grading system, the investigator’s global assessment, and patient satisfaction. A sample was collected from the area that had been treated, and the tissue was examined using hematoxylin and eosin (H–E) staining and immunohistochemistry staining for collagen I. Results: Goodman and Baron global scores showed a statistically significant difference compared to baseline on both sides of the face. However, there was no statistically significant difference between the two treatment modalities. Biopsy specimens showed an increased deposition of collagen I by both laser devices, which was validated and described by immunohistochemistry staining. Conclusion: Both FCO₂ and fractional non-ablative diode lasers proved their efficacy in treating different types of acne scars. For patients who are interested in no downtime and no complications, fractional non-ablative diode laser is recommended as an efficient alternative modality. Full article