The invention provides a composite nano biological glass material and a preparation method thereof. The preparation method comprises the following steps: (1) uniformly mixing ammonia water, absolute ethyl alcohol and deionized water, adding tetraethyl orthosilicate to stir at room temperature, and then, adding triethyl phosphate to continuously stir; (2) adding calcium nitrate terahydrate to stir after ultrasonically dispersing; (3) putting into a drying oven to dry, and putting into a box type electric furnace to treat for 3-4 hours; (4) adding calyx seu fructus physalis extract, polysaccharides in reed rhizome, resveratrol, silk fibroin, hexadecyl trimethyl ammonium bromide, dodecyl dimethyl betaine, sodium alginate and deionized water after cooling, and stirring after ultrasonically dispersing; (5) standing and aging at the room temperature, putting into an ultralow-temperature refrigerator of -70 DEG C for pre-freezing, and performing freeze-drying, thereby obtaining the composite nano biological glass material. The composite nano biological glass material provided by the invention has relatively good mechanical properties, is moderate in air permeability, is beneficial for wound healing, and has very good antibacterial effect.