University of Colorado

Massachusetts Institute of Technology

Other

The General Hospital Corporation

University Technology Corporation

Kristi S Anseth

Christopher Bowman

Charlie Nuttelman

Bilge Hacioglu

Robert Samuel Langer

Theodore W Randolph

Jennifer H Elisseeff

Corinne Lengsfeld

April M Kloxin

Derek Sims

Jennifer L Owens

Mark W Tibbitt

Andrea M Kasko

Mehmet Toner

Venkatram R Shastri

Ajay Shah

Robert S Langer

Balaji Sridhar

Tobin Brown

Brian Polizzotti

Benjamin Fairbanks

Cole DeForest

John Oakey

Blige Hacioglu

Michael D Goodner

Anandkumar R Kannurpatti

Ben Fairbanks

Ian Marozas

Disclosed herein are photodegradable hydrogels and associated kits for selectively capturing and releasing cells. The hydrogels result from cross linking in the presence of a photoinitiator (1) a macromer having a polymeric backbone structure, a photo labile moiety, and a first linking moiety, and (2) a cell-binding moiety having a second linking moiety. These two components are cross-linked by a polymerization reaction of the linking moieties to form a photodegradable hydrogel incorporating the cell-binding moiety within the hydrogel. Also disclosed are methods of making the hydrogels, and methods of using the hydrogels for selectively capturing and releasing cells and for detecting cells in a fluid. Such methods can be used to detect the presence and quantity of certain rare cell types in a biological fluid.

Selective capture and release of rare mammalian cells using photodegradable hydrogels in a microfluidic platform

This invention is in the field of synthesis and amplified photodegradation of hydrogel network and methods of producing and using the same.

Amplified photodegradation of hydrogels and methods of producing the same

A thiol-ene polymeric material is disclosed. The material is produced by the photopolymerization of reactants having thiol and olefin moieties. The material can incorporate encapsulated components, including cells. Additionally, the material can be derivatized by reacting the polymeric material with components such as proteins.

Degradable thiol-ene polymers

A thiol-yne polymeric material and methods for producing said polymers are disclosed. The material is produced by the radically mediated polymerization of monomers having alkyne and thiol functional groups. The alkyne moiety, internal or terminal, may react with one or two thiols. Degradable monomers may be used to form degradable polymers.

Covalently cross linked hydrogels and methods of making and using same

Here, we present a photodegradable microparticle system that can be employed to entrap and deliver bioactive proteins to cells during culture. By using a photosensitive delivery system, experimenters can achieve a wide variety of spatiotemporally regulated release profiles with a single microparticle formulation, thereby enabling one to probe many questions as to how protein presentation can be manipulated to regulate cell function. Photodegradable microparticles were synthesized via inverse suspension polymerization with a mean diameter of 22 &#x3bc;m, and degradation was demonstrated upon exposure to several irradiation conditions. The protein-loaded depots were incorporated into cell cultures and release of bioactive protein was quantified during the photodegradation process. This phototriggered release allowed for the delivery of TGF-&#x3b2;1 to stimulate PE25 cells and for the delivery of fluorescently labeled Annexin V to assay apoptotic 3T3 fibroblasts during culture. By incorporating these photoresponsive protein delivery depots into cell culture, new types of experiments are now possible to test hypotheses about how individual or multiple soluble factors might affect cell function when presented in a uniform, temporally varying, or gradient manner.

Photodegradable groups for tunable polymeric materials

The present invention relates to hydrogels and methods for producing and using the same. In particular, some embodiments of the invention relate to hydrogels and methods for patterning the same.

Hydrogels and methods for producing and using the same

Provided is a method that provides both spatial and temporal control of a polymer degradation process using mono- and multifunctional macromolecular monomers ('macromers') that degrade via single- and multi-photon photolysis mechanisms over a broad range of wavelengths. The macromers can form or be incorporated into networks via covalent, non-covalent and/or ionic interactions. The spatial and temporal degradation of these networks can be controlled. More specifically, provided is a photodegradable macromer, comprising: (a) a photodegradable group; (b) a backbone structure comprising one or more repeating units that may be the same or different, which backbone structure is attached to the photodegradable group directly or through a linker; (c) one or more reactive end groups at one or more ends of the macromer; and optionally, (d) one or more therapeutic agents; and optionally (e) one or more caged groups. Also provided are polymers and networks incorporating macromers of the invention and optionally other substituents such as other polymeric structures. Also provided is a method of controlled degradation of a polymer comprising: providing a photodegradable polymer as described herein and exposing the photodegradable polymer to photoradiation of the appropriate wavelength and energy to cause one or more of the photodegradable groups to photodegrade.

Compositions for tissue engineering and drug delivery have been developed based on solutions of two or more polymers which form semi-interpenetrating or interpenetrating polymer networks upon exposure to active species following injection at a site in a patient in need thereof. The polymers crosslink to themselves but not to each other; semi-interpenetrating networks are formed when only one of the polymers crosslink. The resulting viscous solutions retain the biologically active molecules or cells at the site of injection until release or tissue formation, respectfully, occurs. As a result of studies conducted with polymer-cell suspensions forming interpenetrating polymer networks, it has been determined that polymer solutions can be formulated wherein the active species is provided by exposure of the polymer solution to an exogenous source of active species, typically electromagnetic radiation, preferably light. Studies demonstrate that light will penetrate through skin, body fluids (such as synovial fluid) and membranes and polymerize the polymer solutions. The polymer solutions can be crosslinked ionically or covalently, to form a hydrogel, semi-interpenetrating polymer network or an interpenetrating polymer network.

Semi-interpenetrating or interpenetrating polymer networks for drug delivery and tissue engineering

Methods of forming crosslinked polymer particles in situ from polymer precursors such as monomers or oligomers, comprising exposing a composition comprising at least one polymer precursor, a solvent or solvent mixture, and an antisolvent or antisolvent mixture to photoradiation under conditions whereby particles are formed are provided. The polymer precursor may be photosensitive, or a separate polymerization initiator may be used. In a preferred embodiment, the polymer precursor is insoluble in the antisolvent or antisolvent mixture and the solvent or solvent mixture is soluble in the antisolvent or antisolvent mixture at the concentrations used. Crosslinked polymer particles and crosslinked polymer particles comprising a polymer and a bioactive material are also provided. The polymer may be erodable, and the polymer particles formed may be used in a variety of applications, including controlled release of bioactive materials such as drugs. Polymer particles formed using the methods of the invention have low residual solvent levels and high additive encapsulation efficiencies. The processes of the invention allow control of particle size and morphology, use low operating temperatures and are useful for efficient bulk production.

Preparation and use of photopolymerized microparticles

Methods of forming polymer particles in situ from polymer precursors such as monomers or oligomers, comprising exposing a composition comprising at least one polymer precursor, a solvent or solvent mixture, and an antisolvent or antisolvent mixture to photoradiation under conditions whereby particles are formed are provided. The polymer precursor may be photosensitive, or a separate polymerization initiator may be used. In a preferred embodiment, the polymer precursor is insoluble in the antisolvent or antisolvent mixture and the solvent or solvent mixture is soluble in the antisolvent or antisolvent mixture at the concentrations used. Polymer particles comprising a polymer and a bioactive material are also provided. The polymer may be erodable, and the polymer particles formed may be used in a variety of applications, including controlled release of bioactive materials such as drugs. Polymer particles formed using the methods of the invention have low residual solvent levels and high additive encapsulation efficiencies. The processes of the invention allow control of particle size and morphology, use low operating temperatures and are useful for efficient bulk production.

Growing community of inventors

Boulder, CO, United States of America

Kristi S Anseth