A Method to Reversibly Control Blood Clotting Using Laser Light

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FIG 1. TBA and antidote affect coagulation in whole human blood.  a) Schematic of coronas made from human serum (HS) loaded with NRs and TBA (NR-HS-TBA) + coronas loaded with NBs and antidote (NB-HS-antidote). 800 nm laser irradiation melts the NRs, triggering release of TBA from the coronas, which inhibits thrombin and causes blood coagulation times to increase. Following this, 1100 nm laser irradiation melts the NBs, triggering release of the DNA antidote from the corona. The antidote forms a double-stranded hybrid with TBA, thus restoring thrombin activity and blood coagulation. Fluorescently labeled TBA has a sequence of 5’ GGTTGGTGTGGTTGG-TMR 3’. The fluorescently labeled antidote has the complementary sequence 5’ CCAACCACACCAACC-FAM 3’. Clotting time (tplasma) for a thrombin test using 10 nM thrombin measured by a coagulometer with b) TBA, for c) 500 nM TBA + varying antidote from [anti] = 0 to 1000 nM (anti/TBA = 0 to 2.0).FIG 2. Gold nanoparticles synthesized and loaded for triggered release.  Absorption spectra of a) NRs, NR-HSA-TBA coronas (LSPR max = 777 nm), b) NB, NB-HSA-antidote (LSPR max = 1093 nm). c) TEM image of NRs, scale bar = 20 nm, d) TEM image of NBs, scale bar = 100 nm, e) DH (DLS) of NRs, NR-HS-TBA, NBs, NB-HS-antidote, indicating that a corona contains multiple not a single NR or NB, but multiple ones. f) Zeta potential of NRs, NR-HS-TBA = −9.8 mV, NBs, NB-HS-antidote = −10.1 mV g) Quantified DNA payloads of NR-HS-TBA (674±74 TBA/NR), NB-HS-antidote (1307±255 antidote/NB). h) mixture of NR-CTAB + NB-CTAB before (black) and after (red) 800 nm irradiation. i) NR-CTAB + NB-CTAB before (black) and after (red) 1100 nm irradiation.FIG 3. Release from NR- and NB-coronas and their comparison to covalently loaded NRs.  a) Absorption spectrum of NB-HS-TBA before (black) and after (red) 1100 nm irradiation, where [NB-HS-anti] = 0.3 nM, and released [anti] = 129±5 nM (430±17 anti released/NB). Inset: fluorescence spectrum of released TBA before (black) and after (red) 1100 nm irradiation. b) Absorption spectrum of NR-HS-TBA before (black) and after (red) 800nm irradiation where [NR-HS-TBA] = 2.9 nM, and released [TBA] = 663±23 nM (223±8 DNA released/NR). Inset: fluorescence spectrum of released TBA before (black) and after (red) 800 nm irradiation, c) Effect of the released TBA in blood. Comparing normalized tplasma from released TBA from the coronas [NR-HS-TBA] = 2.9 nM (red), where released [TBA] = 663±23 nM in a clotting test. Supernatant of NR-HS-TBA with exposed to no irradiation and added to blood is defined as tplasma = 1.0 (gray dotted line). A significant difference (p≤0.05) from baseline tplasma is indicated with a * (Table S1), d) tplasma (normalized) calibration curve of free TBA in a thrombin test (stars). Released TBA from NR-HS-TBA (red circle) and extrapolated equivalent concentration (red dashed line). e) tplasma (normalized) calibration curve of free thiolated TBA (blue X’s). Released thiolated TBA from NR-thiol-TBA (1460±108 nM, blue square) and extrapolated equivalent concentration (blue dashed line).
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Inventors
Kimberly Hamad-Schifferli
Lincoln Laboratory, MIT
External Link (web.mit.edu)
Salmaan Baxamusa
Department of Biological Engineering, MIT
Helena De Puig Guixe
Department of Mechanical Engineering, MIT
Managed By
Ben Rockney
MIT Technology Licensing Officer
Patent Protection

Blood Clotting Control

US Patent 9,545,383
Publications
Selective Light-Triggered Release of DNA from Gold Nanorods Switches Blood Clotting On and Off
PLOS ONE, Vol. 8, e68511, 2013

Applications

The methodology can be applied in surgeries, where temporary suspension of blood coagulation is necessary, such as transplants, or on demand for externally controlled rapid halting of bleeding.

Problem Addressed

Treatment of bleeding disorders and surgery blood clotting must be controlled for patient safety. This invention allows reversible control of blood coagulation, using nanoparticles and laser light.  It enables on demand, remote control of blood clotting without the side-effects of commonly used anticoagulants.


Technology

The invention utilizes nanoparticles, which can be introduces into the blood and excited by using laser irradiation to control blood clotting. Nanoparticles, due to their unique properties, can be excited by laser light in a mutually exclusive manner, enabling selective release of a thrombin binding aptamer (TBA), which acts as an anticoagulant to stop blood clotting, or its antidote to revers the effect.

Once the nanoparticles are introduced into the blood stream, a laser at a certain wavelength triggers the release of a TBA from a corresponding nanoparticle when needed. At another wavelength, the laser triggers the release of the TBA antidote, restoring normal blood clotting to the system.

The technology can be usezd to control thrombolysis (clot dissolution) in a similar manner.

Advantages

  • The technology enables on-demand, reversible, remote control of blood clotting.
  • It acts as a substitute for commonly used anticoagulants, often referred to as blood-thinners, without causing negative side-effects due to slow clearance from system and nonspecific targeting.
  • The antidote is specific to the anticoagulant.