Abstract
The use of implants can be hampered by chronic inflammatory reactions, which may result in failure of the implanted device. To prevent such an outcome, the present study examines the anti-inflammatory properties of surface coatings made of either hyaluronic acid (HA) or heparin (Hep) in combination with chitosan (Chi) prepared as multilayers through the layer-by-layer (LbL) technique. The properties of glycosaminoglycan (GAG)-modified surfaces were characterized in terms of surface topography, thickness and wettability. Results showed a higher thickness and hydrophilicity after multilayer formation compared to poly (ethylene imine) control samples. Moreover, multilayers containing either HA or Hep dampened the inflammatory response visible by reduced adhesion, formation of multinucleated giant cells (MNGCs) and IL-1β release, which was studied using THP-1 derived macrophages. Furthermore, investigations regarding the mechanism of anti-inflammatory activity of GAG were focused on nuclear transcription factor-кB (NF-κB)-related signal transduction. Immunofluorescence staining of the p65 subunit of NF-κB and immunoblotting were performed that showed a significant decrease in NF-κB level in macrophages on GAG-based multilayers. Additionally, the association of FITC-labelled GAG was evaluated by confocal laser scanning microscopy and flow cytometry showing that macrophages were able to associate with and take up HA and Hep. Overall, the Hep-based multilayers demonstrated the most suppressive effect making this system most promising to control macrophage activation after implantation of medical devices. The results provide an insight on the anti-inflammatory effects of GAG not only based on their physicochemical properties, but also related to their mechanism of action toward NF-κB signal transduction.
Highlights
Biomaterial implants can trigger an undesired host response upon surgical insertion in the human body leading to limited functionality, longevity and eventually to failure of the biomedical device [1,2]
Characterization of Physical Properties of Coatings The thickness of surface coatings prepared on silicon wafers was studied by ellipsometry in the CharacterizdparriytmiosatnrayteopftooPlyvhe(yreistfhiycylatehlnePedriemoppioneseir)ttio(iPneEsoI)of flpaCoyleyorealttehicanttrgowlsyatsesu.sTedhetompearosuvirdeme eanptsoswiteivree performed for surface charge the for
The polyanionms eHasAureamnendtsHweerpe adnonde ftooridmenatitfiyontheowf ePttEinMg psr.opTehrteiesPoEf Mthes smurafadceecooaftihngysa. lFuigruorne a2Bn and chitosan were desidgenpaictteeddaassigPniEficIa(nHtlAy l/oCwheri)W4HCAAo.fTPhEeMsPcEoMmpsarmedatdo ePEoIfshurefapcaesr.iInn aadndditiconh,iatossigannifiwcanetre abbreviated as PEI(Hep/Chdmiiuf)fl4etriHleanyeceeprs.bceAotmwlpeteoonsgedethtoehf ePtwEr,oI(1Hg0elypsc/oCinshaigm)4lHieneoplga.lyyceanrss wwaesroebsaebrvseodr, bsheodwsinugbtsheeqlouweensttlWy CaAs dfoer scribed in the Materials and Methods Section
Summary
Biomaterial implants can trigger an undesired host response upon surgical insertion in the human body leading to limited functionality, longevity and eventually to failure of the biomedical device [1,2]. It is desirable to design biomaterials that will guide the inflammation process to achieve the desired function of the device. The persistent stimulation of the immune system may shift acute to chronic inflammation, which is characterized by the formation of multinucleated giant cells (MNGCs) as a result of macrophage fusion in an attempt to phagocyte biomaterials larger in size than a single cell [8,9]. Fibroblasts are recruited upon the prolonged inflammatory phase, which may result in encapsulation of the biomedical device causing its failure [10,11]
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