As 3D printing becomes more common and the technique is used to build culture platforms, it is imperative to develop surface treatments for specific responses. The advantages of aminating and oxidizing polystyrene (PS) for human mesenchymal stem cell (hMSC) proliferation and osteogenic differentiation are investigated. We find that ammonia (NH3) plasma incorporates amines while oxygen plasma adds carbonyl and carboxylate groups. Across 2D, 3D, and 3D dynamic culture, we find that the NH3- treated surfaces encouraged cell proliferation. Our results show that the NH3-treated scaffold was the only treatment allowing dynamic proliferation of hMSCs with little evidence of osteogenic differentiation. With osteogenic media, particularly in 3D culture, we find the NH3 treatment encouraged greater and earlier expression of RUNX2 and ALP. The NH3-treated PS scaffolds support hMSC proliferation without spontaneous osteogenic differentiation in static and dynamic culture. This work provides an opportunity for further investigations into shear profiling and coculture within the developed culture system toward developing a bone marrow niche model.