Today's wood protection technology adopts copper-based chemical substances as the widely utilized material. Besides the adverse effect on human health, it also showed negative impact on organisms. To prevent this, a new method using of nano-sized chemical substances in wood preservation was being investigated. In this work, the effects of nano-clay and nano-sized particles of zinc oxide (ZnO), titanium oxide (TiO2), aluminum oxide (Al2O3), and magnesium oxide (MgO) impregnation on the wood samples were investigated. Samples from Scots pine wood were impregnated with the nanoparticles. Besides that, samples with second impregnation of 10% polymethylmethacrylate (PMMA) were also prepared to investigate the improvement in wood properties. The physical and thermal properties of the samples were tested as well as biodegradation against the brown-rot fungus Coniophora puteana. Impregnation of nanoparticles slightly affect the water absorption properties, but second impregnation with 10% PMMA reduced the water absorption up to 20%. The lowest water absorption and the highest water repellencies after 336 h in water were determined on the samples treated with 10% nano-clay-PMMA and MgO-PMMA treatments. Water repellent efficiency and anti-swelling efficiency were also increased by addition of 10% PMMA with little decrement after longer soaking time. The highest ASE was found on the samples treated with 15% of nano-clay and zinc oxide nanoparticles (ZnO) with PMMA. Results also showed that impregnation of nanoparticles provides good protection against the tested fungi with the highest degradation was only 3.88% for Al2O3, compared to control sample with 20.26% weight loss. In conclusion, the applied nanoparticles are suitable substances to increase the durability of pine wood.