Objective: The aim of the study was to compare and evaluate the in vitro biodegradation of nickel and chromium from space maintainers, made of three different companies, i.e (Dantaurum, Rocky mountain and Dtech) in artificial saliva. Materials and methods: The study comprised of 30 space maintainers out of which 10 were fabricated using Dantaurum, 10 using Rocky mountain and 10 using Dtech band materials. Stainless steel wire (Dantaurum, Rocky mountain and Konark) was used for making loops and Leone solder and flux was used for soldering. Each group was further divided into four subgroups containing 1, 2, 3 and 4 space maintainers respectively. The space maintainers in each subgroup were placed in separate glass beakers containing 100 ml of artificial saliva at 37°C for 4 weeks. Salivary samples from each beaker was analyzed for nickel and chromium ions separately on days 1, 7, 14, 21 and 28 days using inductively coupled plasma-optical emission spectrophotometer. Results: Total release of nickel and chromium from all band and loop space maintainers ranged from 0.020 to 1.524 ppm and 0.002 to 0.289 ppm respectively. The release of nickel and chromium between the groups and within the groups was not significant (p < 0.5). Conclusion: There was no substantial release of nickel and chromium from space maintainers made of Dantaurum, Rocky mountain and Dtech which could cause any toxicity. How to cite this article: Anand A, Sharma A, Kumar P, Sandhu M, Sachdeva S, Sachdev V. A Comparative Study of Biodegradation of Nickel and Chromium from Space Maintainers: An in vitro Study. Int J Clin Pediatr Dent 2015; 8(1):37-41.