Abstract
ObjectivesDental implants are major treatment options for restoring teeth loss. Biological processes at the implant‐tissue interface are critical for implant osseointegration. Superior mechanical properties of the implant constitute a major challenge for traditional histological techniques. It is imperative to develop new technique to investigate the implant‐tissue interface.Materials and methodsOur laboratory developed the polyethylene glycol (PEG)‐associated solvent system (PEGASOS) tissue clearing method. By immersing samples into various chemical substances, bones and teeth could be turned to transparent with intact internal structures and endogenous fluorescence being preserved. We combined the PEGASOS tissue clearing method with transgenic mouse line and other labelling technique to investigate the angiogenesis and osteogenesis processes occurring at the implant‐bone interface.ResultsClearing treatment turned tissue highly transparent and implant could be directly visualized without sectioning. Implant, soft/hard tissues and fluorescent labels were simultaneously imaged in decalcified or non‐decalcified mouse mandible samples without disturbing their interfaces. Multi‐channel 3‐dimensional image stacks at high resolution were acquired and quantified. The processes of angiogenesis and osteogenesis surrounding titanium or stainless steel implants were investigated.ConclusionsBoth titanium and stainless steel implants support angiogenesis at comparable levels. Successful osseointegration and calcium precipitation occurred only surrounding titanium, but not stainless steel implants. PEGASOS tissue clearing method provides a novel approach for investigating the interface between implants and hard tissue.
Highlights
Dental and orthopaedic implants have revolutionized the treatment of patients with missing teeth or damaged bones and joints.[1]
We demonstrated that 3‐D imaging based on PEGASOS tissue clearing method is a useful new tool for investigat‐ ing the implant‐tissue interface
The fluorescence preservation is critical for acquiring multi‐channel 3‐D images after tissue clearing and enables applications of multi‐ ple transgenic mouse models to investigate implant‐tissue interface
Summary
Dental and orthopaedic implants have revolutionized the treatment of patients with missing teeth or damaged bones and joints.[1]. Histological sectioning methods remain as the golden standards for visualization and quantitative measurement of peri‐implant angio‐ genesis and new bone formation.[9,13] During the sample preparation, implants were removed to enable tissue embedding and sectioning,[14] which inevitably destroyed the integrity of the interface.[15] Alternatively, ground sections were employed to study implant‐bone interface with‐ out decalcification treatment or removing implants.[15,16] calcein green dynamic labelling and several other histology staining could be performed, the hard tissue embedding process quenched en‐ dogenous fluorescence and compromised most immunofluorescence staining signals.[17,18] In addition, only very few ground sections could be achieved from a sample, which provided very limited information for the whole tissue.[19,20] μCT remains to be the only available approach to investigate an intact bone‐implant interface in 3‐dimension. We demonstrated that 3‐D imaging based on PEGASOS tissue clearing method is a useful new tool for investigat‐ ing the implant‐tissue interface
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