Introduction: Vaginal bleeding during the first trimester of pregnancy, accompanied by a positive fetal heart rate and a closed cervix, is medically referred to as a threatened miscarriage. This condition is considered the most common complication within the first 12 weeks of pregnancy. It is important to note that, this condition is a potential indication of danger to the fetus, and therefore; requires immediate medical attention and monitoring. Objectives: To evaluate the association of putative biomarkers matrix metalloproteinase-9 (MMP-9) and fetal fibronectin (fFN) in threatened miscarriage. Patients and Methods: A case-control study on threatened miscarriage was conducted at Alkarama hospital and Ibn al Balady Hospital in Baghdad. Serum levels of fFN and MMP-9 were analyzed in 91 pregnant women with matching gestational ages, divided into two groups of threatened miscarriage (patient: n=30) and healthy pregnant women (control: n=61). The differences in serum concentration of fFN and MMP-9 between the two groups, as well as their correlation with clinical outcomes, were statistically evaluated. Results: There were variations in serum concentrations of matrix metalloproteinase9 between the threatened miscarriage group (49.34±1.08 pg/mL) and the control group (12.19±4.90 pg/mL). Statistical analysis showed a p-value of 0.009. FFN levels were found to be higher in the threatened miscarriage patient group (467.85±6.22 pg/mL) compared to the control group (230.66±37.44 pg/mL), with a P-value of 0.003. A positive correlation was observed between MMP-9 and fFN (r=0.877, P=0.001). The 95% confidence interval for fFN was (0.9-0.992) and for MMP-9 was (0.78-0.96). Conclusion: Increased levels of MMP-9 and fFN have been associated with a higher risk of fetal demise.