So recall that a difference of squares looks something like this:
[tex] x^{2} -4[/tex] = (x+2)(x-2)
Everything within the parentheses is the exact same-except for the plus and minus signs! If you multiply the first two terms (the two x's in the example above), you'll get [tex] x^{2} [/tex]. If you multiply -2 by x and +2 by x, you'll get -2x + 2x, which equals 0 and cancel out. Then you multiply +2 by -2 to get -4.
So in the above options, only 16[tex] x^{2} [/tex]-81 is a difference of squares, since it simplifies down to (4x + 9)(4x - 9).