Abstract
The use of biopolymers as a three dimensional (3D) support structure for cell growth is a leading tissue engineering approach in regenerative medicine. Achieving consistent cell seeding and uniform cell distribution throughout 3D scaffold culture in vitro is an ongoing challenge. Traditionally, 3D scaffolds are cultured within tissue culture plates to enable reproducible cell seeding and ease of culture media change. In this study, we compared two different well-plates with different surface properties to assess whether seeding efficiencies and cell growth on 3D scaffolds were affected. Cell attachment and growth of murine calvarial osteoblast (MC3T3-E1) cells within a melt-electrospun poly-ε-caprolactone scaffold were assessed when cultured in either “low-adhesive” non-treated or corona discharged-treated well-plates. Increased cell adhesion was observed on the scaffold placed in the surface treated culture plates compared to the scaffold in the non-treated plates 24 h after seeding, although it was not significant. However, higher cell metabolic activity was observed on the bases of all well-plates than on the scaffold, except for day 21, well metabolic activity was higher in the scaffold contained in non-treated plate than the base. These results indicate that there is no advantage in using non-treated plates to improve initial cell seeding in 3D polymeric tissue engineering scaffolds, however non-treated plates may provide an improved metabolic environment for long-term studies.
Highlights
In recent years, the use of biodegradable biopolymers as a structural three-dimensional (3D) support material has become a prominent tissue engineering approach for repairing damaged cartilage or bone [1,2].These biodegradable polymers can be either synthetic or derived from biological sources depending on the intended use [2]
We hypothesize that low adhesive plates would enable a greater cell attachment to the PCL scaffolds owing to cells preferentially attaching to PCL compared to treated tissue culture plates which cells might attach to in preference, leading to lower seeding density on PCL
We demonstrated no significant difference in cell seeding efficiency and proliferation on melt-electrospun PCL tissue engineering scaffolds, cultured in either surface or non-surface treated tissue culture well-plates, over a two week period
Summary
The use of biodegradable biopolymers as a structural three-dimensional (3D) support material has become a prominent tissue engineering approach for repairing damaged cartilage or bone [1,2]. Various manufacturers use chemical and/or physical surface modifications in order to increase cell attachment to their plastic culture plates [13], often through techniques such as corona discharge [14]. We performed an assessment of cell seeding and growth on 3D melt-electrospun tissue engineering poly-ε-caprolactone (PCL) scaffolds cultured in both corona discharge-treated and “low-adhesive” tissue culture well-plates. This was achieved by seeding cells on melt-electrospun PCL scaffolds placed within tissue culture well-plates of each type, and performing periodic DNA quantification and cell metabolism analysis over four weeks. We hypothesize that low adhesive plates would enable a greater cell attachment to the PCL scaffolds owing to cells preferentially attaching to PCL compared to treated tissue culture plates which cells might attach to in preference, leading to lower seeding density on PCL scaffolds in treated tissue plates
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