OBJECTIVE: Although currently available implants can be used to achieve osseointegration under well‐defined conditions, a greater understanding of cell behaviour is required to improve the designs and embark on actual tissue engineering.
MATERIALS AND METHODS: We employed micromachined substrata to investigate some of the main behavioural responses of osteoblasts from rat fetal calvaria to surface topography. In particular, confocal laser scanning microscopy (CLSM), differential interference contrast microscopy, time‐lapse cinemicrography, immunofluo‐rescence, digital radiography and image analysis were used to investigate cell adhesion, cell shape and cytoske‐leton distribution, tissue organization, cell differentiation, and microenvironment.
RESULTS AND CONCLUSIONS A grooved surface permitted the attachment of more cells than a smooth one. Cell shape and cytoskeleton were strikingly influenced as early as 20 min after cell attachment, when the cytoskeleton begins to align with the topography. Some grooved surfaces appeared to promote osteogenesis in vitro as assessed by the production of bone‐like nodules. Moreover, these nodules align with the topography in vitro, and preliminary results indicate that bone‐like tissue also aligns with grooves when such surfaces are implanted in vivo.
Strategies for modifying titanium (Ti) implant surfaces are becoming increasingly popular to enhance osseointegration during acute and inflammatory healing stages. In this study, two dicationic imidazolium-based ionic liquids (IonLs) containing phenylalanine and methionine anions (IonL-Phe(1,10-bis(3methylimidazolium-1-yl)decane diphenylalanine) and IonL-Met-(1,10-bis(3-methylimidazolium-1-yl)decane dimethionine)) were investigated to stably deliver exogenous proteins on Ti to promote osseointegration. The protein selected for this study is High-Mobility Group Box 1 (HMGB1), which recruits inflammatory and mesenchymal stem cells to the implantation site, contributing to healing. To explore IonL−Ti interactions and HMGB1 stability on the IonL-coated surface, experimental characterization techniques including X-ray photoelectron spectroscopy, scanning electron microscopy, dynamic scanning calorimetry (DSC), and liquid chromatography mass spectrometry (LC−MS) were used along with molecular dynamics (MD) computer simulations to provide a detailed molecular level description. Results show well-structured IonL molecules on the Ti surface that impact protein crystallization and coating morphology. IonL cations and anions were found to bind strongly to oppositely charged residues of the protein. LC−MS/MS reveals that HMGB1 B-box lysine residues bind strongly to the IonLs. Stronger interactions of HMGB1 with Ion-Phe in contrast to IonL-Met results in greater retention capacity of HMGB1 in the IonL-Phe coating. Overall, this study provides evidence that the selected IonLs strongly interact with HMGB1, which can be a potential surface treatment for bone-implantable Ti devices.
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