In vitro proliferation and osteogenic differentiation of mesenchymal stem cells on nanoporous alumina

Abstract

Cell adhesion, migration, and proliferation are significantly affected by the surface topography of the substrates on which the cells are cultured. Alumina is one of the most popular implant materials used in orthopedics, but few data are available concerning the cellular responses of mesenchymal stem cells (MSCs) grown on nanoporous structures. MSCs were cultured on smooth alumina substrates and nanoporous alumina substrates to investigate the interaction between surface topographies of nanoporous alumina and cellular behavior. Nanoporous alumina substrates with pore sizes of 20 nm and 100 nm were used to evaluate the effect of pore size on MSCs as measured by proliferation, morphology, expression of integrin β1, and osteogenic differentiation. An MTT assay was used to measure cell viability of MSCs on different substrates, and determined that cell viability decreased with increasing pore size. Scanning electron microscopy was used to investigate the effect of pore size on cell morphology. Extremely elongated cells and prominent cell membrane protrusions were observed in cells cultured on alumina with the larger pore size. The expression of integrin β1 was enhanced in MSCs cultured on porous alumina, revealing that porous alumina substrates were more favorable for cell growth than smooth alumina substrates. Higher levels of osteoblastic differentiation markers such as alkaline phosphatase, osteocalcin, and mineralization were detected in cells cultured on alumina with 100 nm pores compared with cells cultured on alumina with either 20 nm pores or smooth alumina. This work demonstrates that cellular behavior is affected by variation in pore size, providing new insight into the potential application of this novel biocompatible material for the developing field of tissue engineering.

Sika Deer Antler Collagen Type I‐Accelerated Osteogenesis in Bone Marrow Mesenchymal Stem Cells via the Smad Pathway

“…After washing twice with TBST, the membrane was incubated with HRP-labeled secondary antibody (Santa SC-2073) for 1 h at room temperature and washed three times with TBST. Final detection was performed with enhanced chemiluminescence (ECL) Western blotting reagents (Amersham Biosciences, Piscataway, NJ) and membranes were exposed to Lumi-Film Chemiluminescent Detection Film (Roche) [ 27 ]. Loading differences were normalized using a monoclonal β -actin antibody.…”

Biocompatibility of alumina‐based biomaterials–A review

“…Their outcomes revealed that by increasing the pore size of alumina the cell viability and expression of integrin β1 reduced; however, cell morphology stretched, and the ALP activity and mineralization improved. The authors revealed that surface characteristics of alumina have noticeably effects on MG63 osteoblast-like cells responses (Song et al, 2013). However, it has been exhibited that epithelial cells preferred to growth on alumina surfaces with 30 nm scale size in comparison with greater pore sizes (Chung, Son, & Min, 2010).…”

“…Many studied have suggested using surface nanotopography strategy for improving alumina biological functions, although further investigations should be done on defining the effects of using this approach on cells fate over elongated period of time. In addition, more deeply studying the effects of pore sizes of alumina, in a wide range, on the molecular and cell responses should be considered (Song et al, 2013). Although in vitro experiments could help in understanding the general cell responses to the biomaterial, in vivo examinations present better understandings of immunological responses to it.…”

Silk scaffolds with tunable mechanical capability for cell differentiation

“…These results revealed that the scaffolds with good porous structure and nanofibrous topography showed better biocompatibility than the scaffolds from fresh regenerated silk fibroin solution. Some studies have revealed that ECM-mimic nanofibrous topography and porous structures could provide a better microenvironment for cells [30,40,52,53]. Although it is impossible to clarify the specific actions of microporous and nanofibrous structures in improving the biocompatibility, the cell culture results suggested an improved biocompatibility of the scaffolds containing silk nanofibers.…”