Introduction Information on bioavailable Iron (BAI) content in respirable coal dust (RCD) is crucial to address occupational health and safety, especially in preventing coal workers’ pneumoconiosis (CWP). Materials and methods In the present study, we determined BAI concentrations in seventy-seven coal samples collected from ten coal mining regions of Central India. The cytotoxic potential of BAI-RCD was established invitro by using alveolar epithelial (A549) and macrophage (U937) cell lines. The oxidative/antioxidant status, inflammations, and genotoxicity attributed to BAI-RCD exposure were evaluated and correlated with CWP pathophysiology. Results The mean BAI concentrations in the coal samples (n = 77) range from (275 to 9065 mg kg-1) and showed wide variability. Both cell lines were exposed to low (275 mg kg-1), moderate (4650 mg kg-1), and high (9065 mg kg-1) BAI-RCD samples showed significant (p < 0.001) cytotoxicity in a dose-dependent manner (low < moderate < high) compared to the control. After BAI-RCD treatment, both cell lines showed a decrease in antioxidant stress measures (SOD, CAT, and GSH) and a significant (p < 0.001) increase in oxidative stress parameters (NADPH, MPO, LPO, and PC). Furthermore, these cell line models demonstrated a statistically significant (p < 0.001) dose-dependent increase in cytokines (TGF-β1, IL-1β, TNF-α, MCP-1, and IL-6 cytokines) and oxidative DNA damage marker (8-OH-dG). Conclusion Results indicated that the central India coals (even at low BAI content) may be accountable for inflammatory responses and cytotoxicity. Hence, BAI can be important characteristic to establish safety standards for coal dust exposure before active mining.