Can this be used over painted/sealed grout?

I used on sealed grout and it worked great

What is the difference in the pen sizes? Does one contain more product or does it have a bigger tip?

The Large Pen is 15mm wide (about the size of a typical Jumbo Marker). The 5mm pen is about the size of a regular sharpie marker.

What size room does one pen cover?

Good question. The manufacturer states "up to 60 meters coverage for narrow grout joints". 60 meters is approximately 196 linear feet, however results vary as the porosity of grouts can very greatly.

Brand	RAINBOW CHALK MARKERS LTD
Model Number	GP81216
Product Dimensions	3.18 x 3.18 x 15.88 cm; 79 g
Color	ダークグレー
Name List for Name Plates	モダン
Material	着色剤。
Quantity	1
Laser Class	非毒性。
Are Batteries Included	No
Item Weight	79 g

グラウトペン ダークグレー タイルペイントマーカー:防水タイルグラウト着色剤とシーラーペン - ダークグレー、ワイド15mmチップ(20ml)

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グラウトペンダークグレータイルペイントマーカー:防水タイルグラウト着色剤とシーラーペン - ダークグレー、ワイド15mmチップ(20ml)