Enhanced lubrication on tissue and biomaterial surfaces through peptide-mediated binding of hyaluronic acid

Journal name:
Nature Materials
Volume:
13,
Pages:
988–995
Year published:
DOI:
doi:10.1038/nmat4048
Received
Accepted
Published online

Abstract

Lubrication is key for the efficient function of devices and tissues with moving surfaces, such as articulating joints, ocular surfaces and the lungs. Indeed, lubrication dysfunction leads to increased friction and degeneration of these systems. Here, we present a polymer–peptide surface coating platform to non-covalently bind hyaluronic acid (HA), a natural lubricant in the body. Tissue surfaces treated with the HA-binding system exhibited higher lubricity values, and in vivo were able to retain HA in the articular joint and to bind ocular tissue surfaces. Biomaterials-mediated strategies that locally bind and concentrate HA could provide physical and biological benefits when used to treat tissue-lubricating dysfunction and to coat medical devices.

At a glance

Figures

  1. Tissue-surface modification with a HABpep–polymer system.
    Figure 1: Tissue-surface modification with a HABpep–polymer system.

    a, Schematic of a cartilage surface modified with a HABpep designed to interact with and bind HA in surrounding fluid. b, An in vitro covalent strategy for coating the cartilage surface with MAL–PEG–NHS crosslinker, which on reaction with primary amines of the cartilage surface creates an exposed thiol-reactive surface. Subsequently, a thiolated HABpep is reacted to the ​maleimide functionality. On exposure to a HA solution, the HA binds to the peptide–polymer coating on the cartilage surface. c, The PEG crosslinker reaction to articular cartilage was confirmed by ATR-FTIR spectroscopy, which validated the presence of the ether-rich PEG coating with a large ether peak at ~1,066 cm−1. d, PEGylation was further verified by XPS atomic ratios. Compared with unmodified cartilage, coated samples had a carbon to oxygen ratio closer to 2 (the ratio in PEG) and significantly lower nitrogen content. e, HA-binding functionality of the peptide-conjugated cartilage was visualized using a biotinylated HA. Biotinylated HA was synthesized and applied to unmodified cartilage and cartilage modified with the HA-binding polymer system. After thorough washing, the biotinylated HA was treated with streptavidin and horseradish peroxidase for visualization. The tissue surfaces treated with the HA-binding polymer coating stained darker than the untreated native cartilage.

  2. Single-step strategy for the application of the HABpep–polymer system to a tissue surface, and functional translation to a joint environment.
    Figure 2: Single-step strategy for the application of the HABpep–polymer system to a tissue surface, and functional translation to a joint environment.

    a, Schematic of the synthesis of a PEG bifunctional linker with one end group as a HABpep (GAHWQFNALTVR) and another end that either reacts with the amine groups or binds to a tissue surface through an extracellular matrix (ECM)-binding peptide, such as collagen II binding peptide, WYRGRL. First, a HA-binding peptide is linked to a thiol–PEG–SGA linker (i) through an amine–SGA conjugation reaction (ii), followed by the Michael-addition reaction of thiol functionality and ​vinyl dimethyl azlactone (iii). On a tissue surface, this amine-reactive azlactone functionality can be conjugated with either a peptide (iv) that non-covalently binds to ECM components (v), or covalently reacts with the amine functionality present in the tissue (vi). Both (iii) and (iv), with or without HA, can be applied on a tissue surface in a single-step application. b, HA–rhodamine together with HABpep–PEG–Col IIBpep was injected into healthy rat knees in a single step, and HA retention was monitored over time using an IVIS spectrum in vivo imager. HA–rhodamine (white arrows) through the HABpep–polymer system was retained in rat knees even 72 h post-injection, compared with only 6 h without HABpep coating. Scale bar, 1 cm.

  3. Cartilage-surface-bound HA through the HABpep–polymer coating system in the absence of an exogenous lubricant can recapitulate the friction coefficients of high-concentration HA lubricants.
    Figure 3: Cartilage-surface-bound HA through the HABpep–polymer coating system in the absence of an exogenous lubricant can recapitulate the friction coefficients of high-concentration HA lubricants.

    a, Representative schematic for the preparation and incubation of HABpep-coated samples in test solution PBS. Lubrication properties of normal cartilage and severely damaged cartilage coated with the polymer–peptide system were tested in the presence of saline, and compared with uncoated surfaces in either saline or HA. be, Representative graphs of static friction and kinetic friction versus pre-sliding time (s) for the normal cartilage sample (b,c) and severely damaged cartilage sample, osteoarthritis (OA) stage 3–4 (d,e). The error bars indicate mean ± standard deviation; broken lines represent cartilage samples (no HABpep modification) in PBS versus HA bath, and solid lines represent cartilage samples in PBS versus cartilage samples coated with bound HA through HABpep in PBS; one-way analysis of variance Tukey’s test, p ≤ 0.05, n = 3. Cartilage-surface-bound HA through the HABpep–polymer coating system reduced friction values when ​lubricin is extracted from the tissue. The lubrication properties of normal cartilage and severely damaged cartilage (​lubricin removed) coated with the polymer–peptide system were measured in PBS and compared with controls. fi, Representative graphs of static friction and kinetic friction versus pre-sliding time (s) for the normal cartilage sample (f,g) and severely damaged cartilage sample, osteoarthritis stage 3–4 (h,i); the error bars indicate mean ± standard deviation; solid lines represent cartilage samples (​lubricin removed) in PBS versus cartilage samples (​lubricin removed) coated with bound HA through HABpep in PBS; unpaired t-test with the correction method, p ≤ 0.05, n = 3.

  4. Ocular surface application of the HABpep–polymer system.
    Figure 4: Ocular surface application of the HABpep–polymer system.

    a, The HABpep–polymer system as an eye-drop solution can be used to retain HA on the eye surface. Collagen-I-abundant eye tissues without epithelial layers, such as sclera, conjunctiva and cornea, act as anchors for the HABpep–polymer system. b, Fluorescence images of HA retention on untreated and treated eye tissues: sclera, conjunctiva and cornea of untreated eye (i–iii); treated with scrambled collagen I binding peptide Col-IBPep; (YFDEYSLSQS; iv–vi); and treated with collagen I binding peptide (vii–ix). Scale bar, 250 μm. c, Contact lens modification with the HABpep–polymer system was performed by the covalent reaction methodology. d, Fluorescence images for HA–rhodamine retention on a modified contact lens showed relatively darker staining compared with the control. Scale bar, 200 μm.

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Author information

Affiliations

  1. Translational Tissue Engineering Center, Wilmer Eye Institute and Department of Biomedical Engineering, Johns Hopkins University, Baltimore, Maryland 21287, USA

    • Anirudha Singh,
    • Michael Corvelli,
    • Shimon A. Unterman,
    • Peter McDonnell &
    • Jennifer H. Elisseeff
  2. Department of Chemistry, Johns Hopkins University, Baltimore, Maryland 21218, USA

    • Kevin A. Wepasnick

Contributions

The experiments were designed by A.S., M.C., S.A.U., P.M. and J.H.E., and carried out by A.S., M.C., S.A.U. and K.A.W. Data analyses were performed by A.S., M.C. and J.H.E. The manuscript was written by A.S. and J.H.E.

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The authors declare no competing financial interests.

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