Type I collagen with high purity and high denaturation temperature was successfully extracted from the skin of catfish, Pangasianodon hypophthalmus farmed in the Mekong Delta. The effects of the phosphate buffer (PB) concentration, pH, NaCl concentration, incubation temperature of the reaction fluid, and nHA content on the formation of the collagen-nanofibril scaffold were clarified. BP concentration and NaCl concentration had a significant effect on the nanofibril diameter of the scaffold and the mean diameters of the SC nanofibrils are reaching 98 nm and 109 mm corresponding to 90 mM PB and 140 mM NaCl, respectively. The pH also affected the morphology of the scaffold but not the nanofibril diameter of the scaffold. The diameter of collagen fibrils increases with increasing incubation temperature and the collagen fibrils become particle-like at the temperature of 37^oC. nHA content will change the diameter of the collagen fibrils and it is disappeared when nHA to collagen weight ratios are 1/10 and 1/15. Overall results suggested that collagen-nanofibril scaffolds could be prepared in conditions of 60 mM PB, 1 mg mL^-1 collagen, pH 7.4, 93 mM NaCl, the incubation temperature 25 ^oC, and nHA to collagen weight ratio of 1/15.