Chemical Synthesis, Characterisation, and Biocompatibility of Nanometre Scale Porous Anodic Aluminium Oxide Membranes for Use as a Cell Culture Substrate for the Vero Cell Line: A Preliminary Study

Abstract

In this preliminary study we investigate for the first time the biomedical potential of using porous anodic aluminium oxide (AAO) membranes as a cell substrate for culturing the Cercopithecus aethiops (African green monkey) Kidney (Vero) epithelial cell line. One advantage of using the inorganic AAO membrane is the presence of nanometre scale pore channels that allow the exchange of molecules and nutrients across the membrane. The size of the pore channels can be preselected by adjusting the controlling parameters of a temperature controlled two-step anodization process. The cellular interaction and response of the Vero cell line with an in-house synthesised AAO membrane, a commercially available membrane, and a glass control were assessed by investigating cell adhesion, morphology, and proliferation over a 72 h period. The number of viable cells proliferating over the respective membrane surfaces revealed that the locally produced in-house AAO membrane had cells numbers similar to the glass control. The study revealed evidence of focal adhesion sites over the surface of the nanoporous membranes and the penetration of cellular extensions into the pore structure as well. The outcome of the study has revealed that nanometre scale porous AAO membranes have the potential to become practical cell culture scaffold substrates with the capability to enhance adhesion and proliferation of Vero cells.

Biocompatibility of nanometre scale porous anodic aluminium oxide membranes towards the RK 13 epithelial cell line: A preliminary study

“…Detailed surface studies of both membrane types have been carried out by the authors and the reader is referred to these studies for further information. 9,19,20 Cell adhesion studies Optical microscopy and FESEM investigations of RK-13 cells after 24 h of cultivation on all three substrates reveal good cell adhesion. The cells also exhibited a flattened polygonal morphology and wide spread coverage over all three substrate surfaces as seen in (Figure 1).…”

“…The comparative membrane supplied by Whatman ® was found to have a nominal thickness of around 60 µm, a mean pore diameter of 120 ± 45 nm and a mean inter-pore distance of around 0.32 µm. 9,18 The surface topography was also found to be much rougher than the in-house membranes, with numerous jagged edges protruding up from the inter-wall structures between the pores. Detailed surface studies of both membrane types have been carried out by the authors and the reader is referred to these studies for further information.…”

“…2,9 However, for completeness a brief procedure is discussed here. Fabrication begins by cutting up the aluminium sheet into 50 mm x 20 mm strips.…”

“…4 These membranes have also been used for filtration 5 , incorporated into chemical sensors 6,7 and even used as cell scaffolds for potential tissue regenerative procedures. 8,9 From the medical perspective any material being considered as a cell scaffold or substrate needs to take into account biocompatibility and cell-substrate interactions. This is extremely important since these interactions will directly influence cell adhesion, morphology and proliferation.important when you consider molecules involved in cellular processes such as proteins, carbohydrates, nucleic acids and lipids are all nanometre scale structures.…”

Can Anodic Aluminium Oxide Nanomembranes Treated with Nanometre Scale Hydroxyapatite be Used as a Cell Culture Substrate

“…A standard cell culturing procedure used in previous AAO cell line studies by the authors is presented and discussed in reference [29]. All cell-culturing procedures were carried out in triplicate to ensure consistency in the study.…”