Aims and Objectives: To find out any association between Lactate dehydrogenase and Myeloperoxidase enzyme levels in the gingival crevicular fluid at various time interval during alignment of teeth in patients with different levels of crowding. Materials and Method: A total of 20 orthodontic patients were divided into 2 groups (10 patients in each groups) Group A (1-3 mm crowding) Group B (6-9 mm) based on the amount of crowding according to Little’s irregularity index. The GCF sample is collected from the most affected tooth before activation of the appliance (T0), 2 hrs. After activation (T1), 7 days after activation (T2), 14 days after activation (T3) and at the end of alignment phase (T4) using micro capillary pipette. Enzyme quantification was done by ELISA method. Results: Independent sample t- test with unequal variance was done to compare the mean of MPO in minimal and maximum crowding. It was observed that, Maximum crowding displaying statistically significant higher Myeloperoxidase level at 2 hours of time interval compared to minimum crowding. (P =0.028). Similarly the mean of LDH in minimum and maximum crowding was compared. It was observed that, maximum crowding displaying statistically significant higher Myeloperoxidase level at 7th day (P=0.002), at 14th day (P=0.001) and after de-crowding (P=0.002) compared to minimum crowding. Maximum crowding displaying statistically significant higher Lactate dehydrogenase level at 2 hours of time interval compared to minimum crowding. (P =0.001). Contrarily, it was observed that, minimum crowding displaying statistically significant higher Lactate dehydrogenase level at 14 days of time interval compared to maximum crowding. (P =0.002).There is no statistically significant correlation found between Lactate dehydrogenase and Myeloperoxidase in minimum and maximum crowding cases at different time intervals. Conclusion: LDH and MPO activity can be measured with a quick method that is inexpensive and accessible to most laboratories and can be done on chairside with refinement.LDH and MPO activity can rapidly monitor possible deleterious effect of an excessive orthodontic force have been applied, and adjustments can be made according to individual response to orthodontic forces.