The present invention relates to fiber compositions comprising gels or hydrogels. The invention further relates to the composition of a gel or hydrogel loaded biodegradable fiber and methods of fabricating such fibers. The present invention further provides tissue engineering and drug-delivery compositions and methods wherein three-dimensional matrices for growing cells are prepared for in vitro and in vivo use. The invention also relates to methods of manipulating the rate of therapeutic agent release by changing both the biodegradable polymer properties as well as altering the properties of the incorporated gel or hydrogel.

DESCRIPTION OF ILLUSTRATIVE EMBODIMENTS

An embodiment of the invention provides a drug delivery composition comprising at least one fiber, wherein said fiber comprises a first component and a second component, and wherein said first component is a biodegradable polymer and said second component is selected from the group consisting of a gel and a hydrogel. Another embodiment of the invention provides a drug delivery composition comprising a fiber, wherein said fiber comprises a first component and a second component, and wherein said first component is a biodegradable polymer and said second component is water, and further wherein said water is present as an inner core. A further embodiment of the invention provides a drug delivery composition comprising a fiber, wherein said fiber comprises an emulsion consisting essentially of a gel or hydrogel. An embodiment of the invention provides drug delivery composition comprising a fiber, wherein said fiber comprises a first component, and wherein said first component is a gel or hydrogel and further wherein said fiber comprises a hollow bore. An embodiment of the invention provides a scaffold composition comprising one or more fibers, wherein said fibers comprise a first component and a second component, and wherein said first component is a biodegradable polymer and said second component is selected from the group consisting of a gel and a hydrogel. Embodiments of the invention also provide methods of manufacturing the fibers of the present invention.

An embodiment of the invention provides a bi-component fiber where the inner bore of the fiber, i.e., inside diameter of the fiber, comprises a gel or hydrogel and the outer wall of the fiber comprises a biodegradable polymer. As used herein, the term "gel" refers to a colloidal system with at least two phases, one of which forms a continuous three-dimensional network that acts as an elastic solid. As used herein, the term "hydrogel" refers to a colloid in which a dispersed phase (colloid) is combined with a continuous phase (water) to produce a viscous jellylike product.

An alternate embodiment of the invention provides the inverse of the above, i.e. where the outer wall comprises a gel or hydrogel and the inner bore comprises a biodegradable polymer fiber.

Another embodiment of the invention provides a monofilament fiber where a hydrogel or gel is dispersed randomly throughout the biodegradable polymer layer(s). This configuration results in distinct phase separation where the biodegradable polymer fiber constitutes a continuous phase and the gel or hydrogel constitutes a disperse phase. As used herein, a "continuous phase" refers to the liquid in a disperse system in which solids are suspended or droplets of another liquid are dispersed. As used herein, a "disperse phase" refers to the phase of a disperse system consisting of particles or droplets of one system dispersed through another system.

In certain embodiments, where the gel or hydrogel concentration is zero, a water-bored fiber is provided i.e., a fiber in which water is present within the inside diameter of the fiber. In this case, water, optionally in combination with other materials, comprises the inner core of the fiber and the biodegradable polymer fiber comprises the surrounding sheath of the fiber. In an alternate embodiment, the biodegradable polymer fiber sheath comprises a dispersion of gel or hydrogel. In another embodiment, the biodegradable polymer fiber sheath comprises a dispersion of water in place of a dispersion of gel or hydrogel. In other embodiments, the biodegradable polymer fiber sheath comprises a dispersion of water together with a dispersion of gel and hydrogel.

In an embodiment of the invention, the above described fibers are combined with fibers of similar composition. In other embodiments, fibers of dissimilar type and composition are combined.

In an embodiment, a therapeutic agent is incorporated into one or more of the above described fibers, present individually or in combination. In other embodiments, a drug is incorporated into one or more of the above described fibers, present individually or in combination.

In certain embodiments of the invention, a layer of a fiber circumscribes a layer of an adjacent inner fiber. The inner fiber is approximately centered within the outer fiber. In certain embodiments, one or more of the layers of the circumscribed fibers comprise a hydrogel or a gel in the wall of the fiber or in the bore of the fiber. In additional embodiments, a gel or a hydrogel is incorporated as a dispersed phase within the biodegradable polymer of one or more layers of the fibers. Additional embodiments of the invention provide multi-layered fibers, where each layer comprises varying compositions of gels, hydrogels and therapeutic agents. Certain embodiments of the invention provide fibers comprising more than one kind of therapeutic agent within its one or more layers.

The invention further relates to methods of manipulating the rate of therapeutic agent release by changing both the biodegradable polymer properties as well as altering the properties of the incorporated gel or hydrogel. A therapeutic agent-loaded fiber is suitable for implantation in animals, or more preferably in humans as either single strands for use as a therapeutic agent delivery vehicles, or together with other fibers (of either similar or different type) for the formation of a fiber-based scaffold for use in tissue engineering, wound healing, regenerative medicine, or other medically related applications. These fibers may also be used outside the body to create scaffolds for cell culture, tissue culture, or in vitro organogenesis, wherein specific three-dimensional structures of these fibers may be woven, knitted, braided, used as a non-woven mesh, or maintained as parallel, non-parallel, twisted or random arrays for the creation of complex three-dimensional scaffolds. As each fiber within said fiber scaffold might be loaded with different therapeutic agents, and each with a different release kinetics profile, it may be possible to induce specific cell growth into specific regions of the scaffold. This provides the ability to create complicated three-dimensional biological architecture by deliberate placement of specific fibers at specific locations within the fiber scaffold. These three dimensional biological structures may or may not be biomemetic in their design. By the same means, it is possible to release different therapeutic agents to one section of the cell culture, tissue culture, or organoid than to another within the same sample.

This type of complex three-dimensional fiber scaffold may also be implanted into an animal, or a human to induce specific biological responses at different locations within said fiber scaffold. This is accomplished by designing the fiber scaffold such that fibers with specific therapeutic agents and specific release profiles are placed at specific locations within the scaffold. This enables the control of both temporal and spatial therapeutic agent delivery from the fiber scaffold.

"Defined nonhomogeneous pattern" in the context of the current application means the incorporation of specific fibers into a scaffold matrix such that a desired three-dimensional distribution of one or more therapeutic agents within the scaffold matrix is achieved. The distribution of therapeutic agents within the fibers, and possibly within their centers, controls the subsequent spatial distribution within the interstitial medium of the matrix scaffold following release of the agents from the polymer fibers. In this way, the spatial contours of desired concentration gradients can be created within the three dimensional scaffold structure and in the immediate surroundings of the scaffold matrix. Temporal distribution is controlled by the polymer composition and gel or hydrogel composition of the fiber and by the use of multi-layers within a fiber.

One aspect of the present invention is a biocompatible implant composition comprising a scaffold of biodegradable polymer fibers. In various embodiments of the present invention, the distance between the fibers may be about 20 microns, about 70 microns, about 90 microns, about 100 microns, about 120 microns, about 140 microns, about 160 microns, about 180 microns, about 200 microns, about 220 microns, about 240 microns, about 260 microns, about 280 microns, about 300 microns, about 320 microns, about 340 microns, about 360 microns, about 380 microns, about 400 microns, about 450 microns or about 500 microns. In various embodiments the distance between the fibers may be less than 50 microns or greater than 500 microns.

Additionally, it is envisioned that in various embodiments of the invention, the fibers will have a diameter of about 20 microns, about 40 microns, about 60 microns, about 80 microns, about 100 microns, about 120 microns, about 140 microns, about 160 microns, about 180 microns, about 200 microns, about 220 microns, about 240 microns, about 260 microns, about 280 microns, about 300 microns, about 320 microns, about 340 microns, about 360 microns, about 380 microns, about 400 microns, about 450 microns or about 500 microns (including intermediate lengths). In various embodiments the diameter of the fibers may be less than about 20 microns or greater than about 500 microns. Additionally, large fibers with diameters up to 3.5 cm are envisioned for certain embodiments. Preferably, the diameter of the fibers will be from about 60 microns to about 500 microns.

In another embodiment of the present invention, the fibers or a subset of fibers, contain one or more therapeutic agents such that the concentration of the therapeutic agent or agents varies along the longitudinal axis of the fibers or subset of fibers. The concentration of the active agent or agents may vary linearly, exponentially or in any desired fashion, as a function of distance along the longitudinal axis of a fiber. The variation may be monodirectional, that is, the content of one or more therapeutic agents decreases from the first end of the fibers or subset of the fibers to the second end of the fibers or subset of the fibers. The content may also vary in a bidirection fashion, that is, the content of the therapeutic agent or agents increases from the first ends of the fibers or subset of the fibers to a maximum and then decreases towards the second ends of the fibers or subset of the fibers.

In certain embodiments of the present invention, a subset of fibers comprising the scaffold may contain no therapeutic agent. For fibers that contain one or more therapeutic agents, the agent or agents may include: a growth factor, an immunodulator, a compound that promotes angiogenesis, a compound that inhibits angiogenesis, an anti-inflammatory compound, an antibiotic, a cytokine, an anti-coagulation agent, a procoagulation agent, a chemotactic agent, agents that promotes apoptosis, an agent that inhibits apoptosis, a mitogenic agent, a radioactive agent, a contrast agent for imaging studies, a viral vector, a polynucleotide, therapeutic genes, DNA, RNA, a polypeptide, a glycosaminoglycan, a carbohydrate, a glycoprotein. The therapeutic agents may also include those drugs that are to be administered for long-term maintenance to patients such as cardiovascular drugs, including blood pressure, pacing, anti-arrhythmia, beta-blocking drugs, and calcium channel based drugs. Therapeutic agents of the present invention also include anti-tremor and other drugs for epilepsy or other movement disorders. These agents may also include long-term medications such as contraceptives and fertility drugs. They could comprise neurologic agents such as dopamine and related drugs as well as psychological or other behavioral drugs. The therapeutic agents may also include chemical scavengers such as chelators, antioxidants and nutritional agents. Wherein the therapeutic agent promotes angiogenesis, that agent may be vascular endothelial growth factor. The therapeutic agents may be synthetic or natural drugs, proteins, DNA, RNA, or cells (genetically altered or not). As used in the specification and claims, following long-standing patent law practice, the terms "a" and "an," when used in conjunction with the word "comprising" or "including" means one or more.

In general, the present invention contemplates the use of any drug incorporated in the biodegradable polymer fibers of the invention. The word "drug" as used herein is defined as a chemical capable of administration to an organism, which modifies or alters the organism's physiology. More preferably the word "drug" as used herein is defined as any substance intended for use in the treatment or prevention of disease. Drug includes synthetic and naturally occurring toxins and bioaffecting substances as well as recognized pharmaceuticals, such as those listed in "The Physicians Desk Reference," 471st edition, pages 101-321; "Goodman and Gilman's The Pharmacological Basis of Therapeutics" 8th Edition (1990), pages 84-1614 and 1655-1715; and "The United States Pharmacopela, The National Formulary", USP XXII NF XVII (1990), the compounds of these references being herein incorporated by reference. The term "drug" also includes compounds that have the indicated properties that are not yet discovered or available in the U.S. The term "drug" includes pro-active, activated, and metabolized forms of drugs. Tissue stimulating factors are also included such as: dimers of Platelet Derived Growth Factor (PDGF), insulin-like growth factor-1 (IGF-1), IGF-2, basic Fibroblast Growth Factor (bFGF), acidic FGF, Vascular Endothelial Cell Growth Factor (VEGF), Nerve Growth Factor (NGF), Neurotrophic Factor 3 (NT-3), Neurotrophic Factor 4 (NT-4), Brain Derived Neurotrophic Factor (BDNF), Endothelial Growth Factor (EGF), Insulin, Interleukin 1 (II-1), Tumor Necrosis Factor alpha (TNFα.), Connective Tissue Growth Factor (CTGF), Transforming Growth Factor alpha (TGFα), and all other growth factors and cytokines, as well as para-thyroid hormone (PTH), prostaglandin such as Prostaglandin E-1 and Prostaglandin E-2, Macrophage Colony Stimulating Factor (MCSF), and corticosteroids such as dexamethasone, prednisolone, and corticosterone.

The present invention also contemplates the use of hydrogel forming material within the core of the fibers. Hydrogels are structurally stable, synthetic polymer or biopolymer matrices that are highly hydrated. These materials may absorb up to thousands of times their weight in water, (Hoffman, A. S., Advanced Drug delivery Reviews, 43 (2000), 3-12). Hydrogels can be classified into two broad categories: reversible or physical and irreversible or chemical. The networks in physical gels are held together by molecular entanglements and/or secondary forces including ionic, H-bonding or hydrophobic forces. Physical hydrogels are characterized by significant changes in the rheological properties as a function of temperature, ionic concentration, and dilution. Chemical gels, also called permanent gels, are characterized by chemically crosslinked networks. When crosslinked, these gels reach an equilibrium swelling level in aqueous solutions which depends mainly on the crosslink density.

The preparation of hydrogels can be achieved by a variety of methods well known to those of ordinary skill in the art. Physical gels can be formed by: heating or cooling certain polymer solutions (cool agarose, for example), using freeze-thaw cycles to form polymer microcrystals, reducing the solution pH to form a H-bonded gel between two different polymers in the same aqueous solution, mixing solutions of a polyanion and a polycation to form a complex coacervate gel, gelling a polyelectrolyte solution with a multivalent ion of opposite charge, reticulation of linear polymers, grafting of synthetic polymers onto naturally occurring macromolecules, and chelation of polycations (Hoffman, A. S., Advanced Drug delivery Reviews, 43 (2000), 3-12). Chemical gels can be created by crosslinking polymers in the solid state or in solution with radiation, chemical crosslinkers like glutaraldehyde, or multifunctional reactive compounds. They can also be made by copolymerizing a monomer and a crosslinker in solution, copolymerizing a monomer and a multifunctional macromer, polymerizing a monomer within a different solid polymer to form an IPN gel, or chemically converting a hydrophobic polymer to a hydrogel (Hoffman, A. S., Advanced Drug delivery Reviews, 43 (2000), 3-12); Hennick, W. F. and van Nostrum, C. F., Advanced Drug Delivery Reviews, 54 (2002), 13-26.

The present invention contemplates the use of hydrogel precursor materials and non-gelling proteins and polysaccharides within the bore of the fibers. Hydrogel precursor materials are the same materials as those that form hydrogels, but they are not exposed to the agents or conditions that normally gel the materials, or can be other proteins and polysaccharides that form gels but not hydrogels. For example, alginate salts, such as sodium alginate, are gelled in the presence of divalent cations, such as calcium, while other materials create hydrogels via a change in pH or temperature. Certain embodiments of the invention comprise the use of precursor materials that are never gelled. Other embodiments of the invention comprise the use of precursor materials in the fabrication process that later may form gels or hydrogels. The formation of gels or hydrogels in the fiber layer may take place as a part of the fiber fabrication process, after the fiber has been fabricated, or after the application of an appropriate type of external stimuli, including placing the fiber in vitro or in vivo. The terms "gel" or "hydrogel" as used herein is intended to include the formed gel or hydrogel as well as the appropriate precursor molecules involved in the formation of gels and hydrogels.

The biodegradable polymer used for fiber construction may be a single polymer or a co-polymer or blend of polymers and may comprise poly(L-lactic acid), poly(DL-lactic acid), polycaprolactone, poly(glycolic acid), polyanhydride, or natural polymers or polypeptides, such as reconstituted collagen or spider silk and polysaccharides.

The fibers of the claimed invention are manufactured using wet or dry/wet (dry jet wet) spinning. Each method affects the final properties of the fiber being constructed. Wet spinning is a process in which a polymeric material is extruded into a liquid bath containing a coagulant. The coagulant is typically comprised of a non-solvent for the polymer that is miscible with the solvent in the polymer solution, but it can also contain a solvent/non-solvent mixture. In dry jet wet spinning, the polymer solution is first exposed to an air gap before entering the coagulation bath.

In an embodiment of the invention, the fiber comprises a plurality of co-axial layers of biodegradable polymers. The drug delivery fiber of the present invention may be implanted into many sites in the body including dermal tissues, cardiac tissue, soft tissues, nerves, bones, and the eye. Ocular implantation has particular use for treatment of cataracts, diabetically induced proliferative retinopathy and non-proliferative retinopathy, glaucoma, and macular degeneration.

A further aspect of the present invention is a method of producing a fiber-scaffold for preparing an implant capable of controlling the spatial and temporal concentration of one or more therapeutic agents. This method generally comprises forming biodegradable polymer fibers into a three dimensional fiber-scaffold. The biodegradable polymer fibers contain one or more therapeutic agents. The therapeutic agent or agents are distributed in the fiber-scaffold in a defined nonhomogeneous pattern.

In certain embodiments of the invention, gels and hydrogels comprised in the fiber layers may exist at infinitely dilute concentrations, i.e., the concentration of gel or hydrogel is zero, and water is used with or without other substances and/or active agents, including therapeutic agents, in place of the gel or hydrogel.

In one embodiment of this invention, the preferred material for the hydrogel contained in the bore of the fiber will be alginate or modified alginate material. Alginate molecules are comprised of (1-4)-linked β-D-mannuronic acid (M units) and (α-L-guluronic acid (G units) monomers, which vary in proportion and sequential distribution along the polymer chain. Alginate polysaccharides are polyelectrolyte systems that have a strong affinity for divalent cations (e.g. Ca²⁺, Sr²⁺, Ba²⁺) and form stable hydrogels when exposed to these molecules. The biodegradable polymer is poly(L-lactic acid) (PLLA). In an embodiment, the alginate is contained as the inner core and the PLLA is the outer sheath. The concentration of alginate is in the range of 0.25 w/v % to 100 w/v % (i.e., g/100 ml water), preferably in the range of 0.75 w/v % to 20 w/v %, and most preferably at a concentration of 1 w/v %. The source and composition of alginate directly affects its usable concentration.

In another embodiment of this invention, the PLLA sheath surrounding the inner gel or hydrogel core comprises a cocktail of PLLA polymers of different molecular weights as a means of increasing the degradation rate. The proportions of the PLLA polymers and the range of the polymer molecular weights can vary. In an exemplary embodiment, the polymer cocktail comprises 80% by weight of a PLLA polymer of Mw=100,000 Daltons; 15% by weight of a polymer of Mw=2,000 Daltons; and 5% by weight of a polymer Mw=300,000 Daltons.

In another embodiment of the invention, the PLLA sheath surrounding the inner gel or hydrogel core is comprised of two phases, a continuous phase comprising a biodegradable polymer and a dispersed phase comprising an aqueous phase stabilized by a surfactant. The aqueous phase may optionally comprise therapeutic agents. The amount of the dispersed phase ranges from about 0% to about 85% by weight relative to the weight of the fiber. In a preferred embodiment the amount of the dispersed phase ranges from about 33% to about 50% by weight relative to the weight of the fiber. As the ratio of the dispersed phase increases, so does the rate of degradation of the polymer. This leads to increased release rates of loaded therapeutic agents.

In an embodiment of this invention, agents that are designed to degrade the gel or hydrogel are loaded into the dispersed aqueous phase of the biodegradable polymer component of the fiber (as described above). This agent is released into the gel or hydrogel slowly over time to break down the gel or hydrogel. This increases therapeutic agent release rates. In addition, many of the potential gels and hydrogels are not directly biodegradable within animals, or more especially humans. Therefore, this planned degradation helps the body to eliminate the gels or hydrogels when they are no longer needed.

In an embodiment, the alginate is gelled internally by the addition of gelling agents added directly to the alginate solution. Typical gelling agents include calcium chloride, calcium carbonate, calcium-EDTA (Ethylene Diamine Tetracetic Acid), or other compounds containing bivalent cations that are well known to those skilled in the art. The concentration of the gelation agent ranges from about 5 mM to about 100 mM, more preferably from about 12 mM to about 50 mM, and most preferably from about 15 mM to 30 mM. The range chosen is determined by desired hydrogel properties. If not readily soluble at neutral pH, the gelling agent is typically activated by a drop in pH of the solution. This acidification can be achieved through a number of acids or lactones. This list includes, but is not limited to, citric acid, hydrochloric acid, D-glucono-delta-lactone, and glacial acetic acid.

In another embodiment, the gel or hydrogel is gelled externally by incorporating the gelling agent source into the biodegradable fiber. Alternately, the gelling agent source is added to a water phase that is loaded into one or more layers of the biodegradable polymer. In this way, the gelling agent is slowly released into the gel or hydrogel as the fiber degrades. In certain embodiments, as the fiber degrades and becomes weaker and more porous, the gel becomes more tightly cross-linked. In this way, it may be possible to continuously alter the release rate as the fiber degrades. Release rates tend to increase as the polymer becomes more porous, in this case, this trend would be offset by the gel becoming more tightly cross-linked, hence retarding release rates through the gel or hydrogel as the fiber degrades.

In another embodiment, the gelling agent is soluble in the polymer solvent and is mixed with the polymer solution at the time of fiber fabrication. In this embodiment, rather than the gelling agent being maintained in an aqueous phase, it is molecularly mixed with the polymer. The same net effect of releasing the gelling agent into the gel or hydrogel slowly as the fiber degrades. This embodiment allows the use of organically soluble sources of gelling agents.

In another embodiment, the gelation agents are carried within the alginate solution that are activated over time, such as within lipospheres, microspheres, nanoparticles or other encapsulants that are activated later. These may be slowly activated over time, or purposefully activated by some external event. This will result in the gel either being strengthened, or maintained over time.

In another embodiment of the invention, the gel or hydrogel is the exterior sheath and the biodegradable polymer is the interior core. In this embodiment the gelling agent is in the coagulating bath, which would be an external gelation.

The present invention provides compositions and methods to create single, drug releasing fibers as well as the composition and methods to create a heterogeneous, woven, knitted, braided, non-woven, twisted, parallel array or random three-dimensional fiber scaffold for growing cells in tissue engineering applications. These scaffolds can be used in vitro and in vivo, and due to their heterogeneity can create both spatial and temporal distributions of therapeutic agents. In this invention, therapeutic agents may include drugs, proteins, peptides, mono- and di-saccharides, polysaccharides, glycoproteins, DNA, RNA, viruses, or other biological molecules of interest. The term therapeutic agent in this invention also includes radioactive materials used to help destroy harmful tissues such as tumors in the local area, or to inhibit growth of healthy tissues, such as in current stent applications; or markers to be used in imaging studies.

A. Three Dimensional Fiber Scaffolds

To create the heterogeneous scaffolds of the present invention, the therapeutic agents are encapsulated into individual fibers of the matrix by methods to be described herein. The therapeutic agents are released from each individual fiber slowly, and in a controlled manner. The fiber format has many advantages as a drug delivery platform over other slow drug-releasing agents known to those familiar in the art such as microspheres, porous plugs or patches. The primary advantage of fibers is that they can provide complex three-dimensional woven, or non-woven scaffolding, with or without patterning, to allow cells to attach, spread, differentiate, and mature into appropriately functioning cells. Because they can form patterns, a "smart scaffold" can be produced to induce cells of specific types to migrate to specific regions of the scaffold due to specific chemotactic factors being released. This scaffold mimics the function of the extracellular matrix material both during embryological development and in post-embryological tissues. Additionally, filaments could be formed into a unique scaffold that provides a growth substrate for tissue repair or reconstruction that is not reminiscent of a natural like structure.

Because of the ability to weave patterns to induce appropriate cell types into specific regions, it is possible to incorporate strands that will induce the formation of blood vessels into the fabric. This may be accomplished by providing fibers that release growth factors such as vascular endothelial growth factor (VEGF). By appropriate spacing of VEGF containing-fibers into the weave pattern, large tissues may be engineered, and the cells in such tissues can be provided with a sufficient blood supply and thereby receive oxygen and nutrients and enable the removal of waste products.

Fibers also have the advantage of providing the body with short term mechanical support in such applications as stents, wherein the polymer fiber can maintain the lumen of any tubular body, such as arteries, veins, ducts (e.g. bile duct, ureter, urethra, trachea, etc.), organs of the digestive track such as esophagus, intestine, colon, and connective tissue such as tendons, ligaments, muscle and bone. The fibers provide a useful structure to support mechanical strength or tension during the healing process. Fibers may also be useful to promote neural regeneration or reconstruction of nerves or spinal cord.

B. Fiber Formats

There are a large number of combinations and variations within the scope of this invention. This invention covers gel or hydrogel combinations with a biodegradable polymer fiber in a multi-layer, multi-component format, where each layer is fully contained within the next outer layer, and the inner layer is generally centered within the outer layer. These layers can be comprised of different gels or hydrogels, or different biodegradable polymers.

This invention also includes the use of gels or hydrogels as a dispersed phase within biodegradable polymer layer, wherein the continuous phase is the biodegradable polymer phase. The dispersed phase may be stabilized by either an internal or external surfactant.

In the case of the dispersed gel or hydrogel within the biodegradable polymer layer, and in the case of the gel or hydrogel layer being interior to a biodegradable polymer layer, an allowable special case is that the concentration of the hydrogel is zero. This means that water may be used (with or without the inclusion of other substances) in the place of the gel or hydrogel.

As an additional special case, it may be possible for the polymer concentration in the innermost core to be zero, in which case the solvent normally used with the polymer is replaced by a non-solvent. In this case, the non-solvent core acts as an internal coagulating bath. The result is that a hollow fiber is created. This special case can occur with or without a gel or hydrogel exterior to the biodegradable polymer layer(s) and with or without a dispersed gel, hydrogel or water phase within the biodegradable polymer layer(s).

This leads to a large number of potential combinations. The basic types are external biodegradable polymer with internal gel or hydrogel, and the inverse design, i.e. gel or hydrogel external with the biodegradable polymer as the internal core. In each of these combinations, the biodegradable polymer layer may or may not have a dispersed water, gel or hydrogel phase. Another case is a monofilament fiber with a gel or hydrogel dispersed phase.

C. Release Kinetics of Individual Fibers

Further, there are various means for controlling the release kinetics of the therapeutic agent, thus temporally controlling the release of the therapeutic agent. The following discussion will pertain only to the fiber format wherein the polymer sheath surrounds an inner core of gel or hydrogel. The first point of control for the polymer is to mix low molecular weight polymer in with the higher molecular weight, fiber forming polymers. In this way, the lower molecular weight component is able to rapidly degrade and diffuse from the fiber, making the fiber more porous. This makes the interior therapeutic agents within the gel or hydrogel more accessible. A second means of accelerating the release rate of the fiber is to create a bi-phasic fiber, wherein the continuous phase is the biodegradable polymer, and the dispersed phase is aqueous pockets that are stabilized by a surfactant. As the concentration of the dispersed phase increases, a pathway is created from the outside to the inner gel or hydrogel where the only polymer that must be degraded is between the various pockets of the dispersed aqueous phase. This has the effect of leaving much less polymer to degrade to connect the gel or hydrogel to the outside world, thus accelerating the release of the therapeutic agent. It is also possible for this dispersed aqueous phase to contain the same or a different drug or therapeutic agent. In this case, the drug or therapeutic agent in the dispersed aqueous phase will be released first, followed by the release of the therapeutic agent in the gel or hydrogel. To alter the release kinetics of the drug or therapeutic agent in the polymer fiber wall, it is possible to slightly adapt the above description such that the dispersed phase is now a gel or hydrogel as opposed to being aqueous. In this case, the fluid pathway shortening exists as in the case of an aqueous dispersed phase; however, the connecting pathway must now go through pockets of gel or hydrogel, wherein the diffusion of the therapeutic agent is retarded compared to a purely aqueous pathway. The degree to which the diffusion is retarded is a function of the type of gel or hydrogel, the type and degree of cross-linking, and the concentration of the gel or hydrogel. All of these parameters are within the control of the entity forming the fiber. It is also possible to control the concentration of the dispersed aqueous or gel phase within the biodegradable polymer as a function of distance along the long axis of the fiber. By this means, it is possible to have different release kinetics at one end of the fiber than at the other, with a defined gradient of release kinetics down the length of the fiber. This change in release kinetics may or may not be combined with a gradient of therapeutic agent concentration. By the same means, it is possible to have the content of the disperse phase vary as a function of distance down the polymer fiber such that at one end the dispersed phase would be for example purely aqueous and at the second end of the fiber, the dispersed phase could be a gel or hydrogel. Other gradients are also possible including varying concentrations of the gel within the disperse phase. Thus a great deal of control is available on the release kinetics of the fiber. Aside from these changes in the polymer wall of the fiber, it is also possible to control the release kinetics from this fiber by altering the type, concentration, and degree of cross-linking within the gel or hydrogel in the core of the fiber, which contains a therapeutic agent.

The ability to dynamically change the release kinetics of the gel or hydrogel being loaded into the core or as a dispersed phase within a biodegradable polymer fiber over the course of the drug delivery period constitutes an important aspect of the invention. This affords unique opportunities that are not possible to be present in other forms of drug delivery from gels or hydrogels. The first means of control available because of the gel being loaded into a biodegradable polymer fiber is the ability of this fiber to release agents known to cross link the gel. In this way, over time, the cross-linking density of the gel actually increases, which will retard the release of the therapeutic agent. This release of the cross linking agent from the biodegradable polymer fiber sheath is itself controllable by means outlined above, i.e. using a cocktail of molecular weights, or changing the concentration of the dispersed aqueous phase. As a special case of the biodegradable polymer fiber sheath is a multi-layer, and multi-component biodegradable polymer sheath. This allows the creation of directional specificity, as well as changes in the release kinetics from each layer of the biodegradable polymer fiber sheath. For example, consider the case of two layers of biodegradable polymer fiber in the sheath. The innermost layer could contain agents that act to cross link the gel or hydrogel core of the fiber, and this layer could be composed of a biodegradable polymer that has a rapid degradation rate. Further, this layer could contain a high degree of dispersed aqueous phase. In this same example, the outermost layer may be composed of a different biodegradable polymer with a different degradation rate, and a different concentration of dispersed aqueous (or gel or hydrogel) dispersed phase, including zero. This example would create a situation where the cross-linking agent would be delivered inwardly to the gel or hydrogel in the core of the fiber over time, thus creating a situation wherein the diffusion coefficient of the therapeutic agent loaded into the gel or hydrogel in the core of the fiber decreases over time.

Another special case is where the polymer fiber contains agents that degrade the gel or hydrogel in the core of the fiber. Using the same logic as explained above, this too creates a situation where the diffusion coefficient of the therapeutic agent in the gel or hydrogel in the core or dispersed within the fiber changes continuously over time. In this case, however, the diffusion rate increases over time. This particular case also has the advantage that the body of the animal or preferably the human into which the fiber is implanted may not have the specific enzymes or other chemical conditions required to degrade the gel or hydrogel. In this case, loading appropriate degradation agents into the wall of the fiber allows the degradation of the gel or hydrogel, and thus aids the clearance of the gel or hydrogel from the host. Again, as described above, the release of the degradation agents is largely controllable by changing properties of the biodegradable polymer layers in the sheath of the fiber.

By these methods, it is seen that the release kinetics of the therapeutic agent from a gel or hydrogel core or dispersed in a sheath of biodegradable polymer fiber is alterable by virtue of the presence of biodegradable polymer sheath.

In the case where the gel or hydrogel is the exterior layer and the biodegradable polymer is the core of the fiber. In this case the biodegradable polymer core may consist of one or more multi-component layers as described above, and again each layer may contain a different concentration of dispersed aqueous or gel or hydrogel phase, which may or may not themselves carry therapeutic agents. The overall release of therapeutic agent(s) from the fiber is controlled by the location of the therapeutic agents, either in the gel or hydrogel exterior, or within the biodegradable polymer core or both. By the same means as described above, the exterior gel or hydrogel release kinetics may be altered by the release of cross-linking, or degrading agents from the biodegradable polymer fiber core. As these agents are released from the biodegradable polymer fiber core, they will alter the properties of the exterior gel or hydrogel, thus decreasing or increasing the diffusion of the therapeutic agent from the exterior gel or hydrogel. For any therapeutic agent(s) within the biodegradable polymer core, the release of these agents is controlled on two levels. First, as explained above the type and molecular weight distribution of the polymer itself changes the release kinetics, as well known to those skilled in the art. In addition to this, the concentration of any dispersed aqueous or gel or hydrogel phase will alter the release from the biodegradable polymer. However, as the gel or hydrogel is surrounding the biodegradable fiber, all therapeutic agents within the biodegradable polymer must diffuse through the gel or hydrogel. Therefore, any changes to the diffusion of the therapeutic agent(s) through the gel or hydrogel also directly affect the release of any therapeutic agents within the core of the fiber. Therefore, in this case, one can change the release kinetics of the fiber by altering both the gel and the biodegradable polymer segments.

If the dispersed phase is a gel or hydrogel that also contains the therapeutic agent, then the release of that therapeutic agent is controllable by the same means of choice of biodegradable polymer, molecular weight distribution, and concentration of the dispersed phase. In addition, the properties of the gel or hydrogel also alter the release of the therapeutic agent from the dispersed phase within the monofilament fiber.

D. Biodegradable Polymers

Preferred polymers for use in the present invention include single polymer, co-polymer or a blend of polymers of poly(L-lactic acid), poly(DL-lactic acid), polycaprolactone, poly(glycolic acid) or polyanhydride. Naturally occurring polymers may also be used such as reconstituted collagen or natural silks. Those of skill in the art will understand that these polymers are just examples of a class of biodegradable polymer matrices that may be used in this invention. Further biodegradable matrices include polyanhydrides, polyorthoesters, and poly(amino acids) (Peppas and Langer, 1994). Any such matrix may be utilized to fabricate a biodegradable polymer matrix with controlled properties for use in this invention. A non-exhaustive list of biodegradable polymers that produce non-toxic degradation products are listed in Table 1.

E. Types of Gels and Hydrogels

In simple terms, a gel is a liquid system that acts like a solid. More technically defined, a gel is a colloidal system with at least two phases, one of which forms a continuous three-dimensional network that acts as an elastic solid. Gel formation through physical, molecular, or chemical association results in an infinite molecular weight for the system. The viscoelastic material formed has a storage modulus, G′, that is greater than the loss modulus, G", and both G′ and G" are almost independent of frequency. [E. R. Morris, Polysaccharide solution properties: origin, rheological characterization and implications for food systems, Frontiers in Carbohydrate Research 1: Food Applications (R. P. Millane, J. N. BeMiller, and R. Chandrasekaran, eds.), Elsevier, London, 1989, p. 132.] The storage modulus characterizes the rigidity of the sample, while the loss modulus characterizes the resistance of the sample to flow. [Damodaran, Srinivasan, Food Proteins and Their Applications, Food Science and Technology (Marcel Dekker, Inc.); New York Marcel Dekker, Inc., 1997.] Examples are polymer solutions, micellar solutions, microemulsions and, in more recent years, the field has been extended with the large number of organic solvents that are gelled by the presence of small organic molecules at very low concentrations.

A hydrogel is defined as a colloid in which the disperse phase (the colloid) has combined with the continuous phase (water) to produce a viscous jellylike product. [Dictionary of Chemical Terms, 4th Ed., McGraw Hill (1989)]. Hydrogels are able to swell rapidly in excess water and retain large volumes of water in their swollen structures. The polymeric material comprising the hydrogel can absorb more than 20% of its weight in water, though formed hydrogels are insoluble in water and they maintain three-dimensional networks. [Amidon, Gordon L., Transport Processes in Pharmaceutical Systems, Drugs and the Pharmaceutical Sciences; v. 102 New York Marcel Dekker, Inc., 2000]. They are usually made of hydrophilic polymer molecules crosslinked either by chemical bonds or by other cohesion forces such as ionic interaction, hydrogen bonding, or hydrophobic interaction. [J. I. Kroschwitz, Concise Encyclopedia of Polymer Science and Engineering, New York, Wiley, XXIX, p 1341, 1990.]

Hydrogels are -elastic solids in the sense that there exists a remembered reference configuration to which the system returns even after being deformed for a very long time.

An organogel is defined as an organic phase with an interlaced polymeric component. Preferred solvents include non-toxic organic solvents including, but not limited to, dimethyl sulfoxide (DMSO), mineral oils and vegetable oils. The term "organogel" was initially used to describe a specific concept of gelation, by a gelatin solution, of a water-in-oil inverse microemulsion (see Luisi et al. Colloid & Polymer Science, 1990, vol. 268, p. 356-374). The term has recently been extended to gelled systems comprising two immiscible phases (water in oil) stabilized in lecithin enriched with phosphatidylcholine and usually hydrogenated (see Williman et al. Journal of Pharmaceutical Sciences, 1992, vol. 81, p. 871-874, and Schchipunov et al., Colloid Journal, 1995, vol. 57, p. 556-560). These emulsions have a lamellar phase and are in the form of gels even in the absence of gelling agents, hence the name organogels, which denotes this type of emulsion irrespective of the orientation of the emulsion (Water-in-Oil or Oil-in-Water).

The types of gel materials used in the present invention include polysaccharides, including but not be limited to, amylose, amylopectin, glycogen, cellulose, hyaluronate, chondroitin, heparin, dextrin, inulin, mannan, chitin, galactose, guar gum, carrageenan, agar, furcellaran, xanthan gum, other hydrocolloid gums, pectin, locust bean gum, acacia, ghatti gum, pentosan, arabinogalactan, synthetic derivatives thereof, and mixtures thereof

Examples of materials which can form hydrogels include natural and synthetic polysaccharides and other natural and synthetic polymers and their derivatives, and combinations of these. Suitable polysaccharides and polymers include but are not limited to: amylose, amylopectin, glycogen, cellulose, hyaluronic acid, chondroitin sulfate, heparin, dextrin, inulin, mannan, chitin, galactose, guar gum, carrageenan, agar, furcellaran, xanthan gum, other hydrocolloid gums, pectic acid and pectin, locust bean gum, acacia, ghatti gum, pentosan, arabinogalactan, alginates and alginate derivatives, gellan, gellan gum, glucose, collagen (and gelatin), cellulose, carboxymethylcellulose, hydroxymethylcellulose, hydroxypropylmethylcellulose, methylcellulose, and methoxycellulose, fibrin, xanthan and xanthan gum, agarose, chitosan (polycationic polysaccharide polymers), albumin, human gamma globulin, pullulan, carrageenan (polyanionic polysaccharide polymers), dextrin, dextran, dextran sulfate, keratin, inulin, dextrose, amylose, glycogen, amylopectin, polylysine and other polyamino acids, polyesters such as polyhydroxybutyrate and polyphosphazines, poly(vinyl alcohols), poly(alkylene oxides) particularly poly(ethylene oxides), polyethylene glycol (including PEO-PPO-PEO and the like block copolymers like Pluronics®), poly(allylamines) (PAM), poly(acrylates), modified styrene polymers, pluronic polyols, polyoxamers, polypropylenes, polyurethanes, poly(uronic acids), polyvinyl chloride, poly(vinylpyrrolidone) and copolymers, graft copolymers, synthetic derivatives, blends and other mixtures of the above. Polysaccharides are the preferred polymers for this invention. Alginate, for example, is biocompatible, non-cytotoxic, non-carcinogenic, non-inflammatory, and non-immunogenic, and, therefore, a good candidate for use.

F. Types of Polymeric Materials

Exemplary natural polymers include naturally occurring polysaccharides, such as, for example, arabinans, fructans, fucans, galactans, galacturonans, glucans, mannans, xylans (such as, for example, inulin), levan, fucoidan, carrageenan, galatocarolose, pectic acid, pectins, including amylose, pullulan, glycogen, amylopectin, cellulose, dextran, dextrin, dextrose, glucose, polyglucose, polydextrose, pustulan, chitin, agarose, keratin, chondroitin, dermatan, hyaluronic acid, alginic acid, xanthan gum, starch and various other natural homopolymer or heteropolymers, such as those containing one or more of the following aldoses, ketoses, acids or amines: erythrose, threose, ribose, arabinose, xylose, lyxose, allose, altrose, glucose, dextrose, mannose, gulose, idose, galactose, talose, erythrulose, ribulose, xylulose, psicose, fructose, sorbose, tagatose, mannitol, sorbitol, lactose, sucrose, trehalose, maltose, cellobiose, glycine, serine, threonine, cysteine, tyrosine, asparagine, glutamine, aspartic acid, glutamic acid, lysine, arginine, histidine, glucuronic acid, gluconic acid, glucaric acid, galacturonic acid, mannuronic acid, glucosamine, galactosamine, and neuraminic acid, and naturally occurring derivatives thereof Accordingly, suitable polymers include, for example, proteins, such as albumin.

Exemplary semi-synthetic polymers include carboxymethylcellulose, hydroxymethylcellulose, hydroxypropylmethylcellulose, methylcellulose, and methoxycellulose. Exemplary synthetic polymers include polyphosphazenes, polyethylenes (such as, for example, polyethylene glycol (including the class of compounds referred to as Pluronics®, commercially available from BASF, Parsippany, N.J.), polyoxyethylene, and polyethylene terephthlate), polypropylenes (such as, for example, polypropylene glycol), polyurethanes, polyvinyl alcohol (PVA), polyvinyl chloride and polyvinylpyrrolidone, polyamides including nylon, polystyrene, polylactic acids, fluorinated hydrocarbon polymers, fluorinated carbon polymers (such as, for example, polytetrafluoroethylene), acrylate, methacrylate, and polymethylmethacrylate, and derivatives thereof.

The polymeric materials are selected from those materials which can be polymerized or their viscosity altered in vivo by application of exogenous means, for example, by application of light, ultrasound, radiation, or chelation, alone or in the presence of added catalyst, or by endogenous means, for example, a change to physiological pH, diffusion of calcium ions (alginate) or borate ions (polyvinyl alcohol) into the polymer, or change in temperature to body temperature (37° C.).

G. Agents that Promote Angiogenesis

One class of therapeutic agents to be encapsulated by the polymer fibers of the present invention are therapeutic agents that promote angiogenesis. The successful engineering of new tissue requires the establishment of a vascular network. The induction of angiogenesis is mediated by a variety of factors, any of which may be used in conjunction with the present invention (Folkman and Klagsbrun, 1987, and references cited therein, each incorporated herein in their entirety by reference). Examples of angiogenic factors includes, but is not limited to: vascular endothelial growth factor (VEGF) or vascular permeability factor (VPF); members of the fibroblast growth factor family, including acidic fibroblast growth factor (aFGF) and basic fibroblast growth factor (bFGF); interleukin-8 (IL-8); epidermal growth factor (EGF); platelet-derived growth factor (PDGF) or platelet-derived endothelial cell growth factor (PD-ECGF); transforming growth factors alpha and beta (TGF-α, TGF-β); tumor necrosis factor alpha (TNF-α); hepatocyte growth factor (HGF); granulocyte-macrophage colony stimulating factor (GM-CSF); insulin growth factor-1 (IGF-1); angiogenin; angiotropin; angiotensin; fibrin and nicotinamide (Folkman, 1986, 1995; Auerbach and Auerbach, 1994; Fidler and Ellis, 1994; Folkman and Klagsbrun, 1987; Nagy et al., 1995).

H. Cytokines

In certain embodiments the use of particular cytokines incorporated in the polymer fibers of the present invention is contemplated. Table 2 below is an exemplary, but not limiting, list of cytokines and related factors contemplated for use in the present invention.

G. Polynucelotides

The polynucleotides to be incorporated within the polymer fibers of the present invention extend to the full variety of nucleic acid molecules. The nucleic acids thus include genomic DNA, cDNAs, single stranded DNA, double stranded DNA, triple stranded DNA, oligonucleotides, Z-DNA, mRNA, tRNA and other RNAs. DNA molecules are generally preferred, even where the DNA is used to express a therapeutic RNA, such as a ribozyme or antisense RNA.

A "gene" or DNA segment encoding a selected protein or RNA, generally refers to a DNA segment that contains sequences encoding the selected protein or RNA, but is isolated away from, or purified free from, total genomic DNA of the species from which the DNA is obtained. Included within the terms "gene" and "DNA segment", are DNA segments and smaller fragments of such segments, and also recombinant vectors, including, for example, plasmids, cosmids, phage, retroviruses, adenoviruses, and the like.

The term "gene" is used for simplicity to refer to a functional protein or peptide encoding unit. As will be understood by those in the art, this functional term includes both genomic sequences and cDNA sequences. "Isolated substantially away from other coding sequences" means that the gene of interest forms the significant part of the coding region of the DNA segment, and that the DNA segment does not contain large portions of naturally-occurring coding DNA, such as large chromosomal fragments or other functional genes or cDNA coding regions. Of course, this refers to the DNA segment as originally isolated, and does not exclude genes or coding regions, such as sequences encoding leader peptides or targeting sequences, later added to the segment by the hand of man.

The present invention does not require that highly purified DNA or vectors be used, so long as any coding segment employed encodes a selected protein or RNA and does not include any coding or regulatory sequences that would have a significant adverse effect on the target cells. Therefore, it will also be understood that useful nucleic acid sequences may include additional residues, such as additional non-coding sequences flanking either of the 5′ or 3′ portions of the coding region or may include various internal sequences, i.e., introns, that are known to occur within genes.

Many suitable DNA segments may be obtained from existing, including commercial sources. One may also obtain a new DNA segment encoding a protein of interest using any one or more of a variety of molecular biological techniques generally known to those skilled in the art. For example, cDNA or genomic libraries may be screened using primers or probes with designed sequences. Polymerase chain reaction (PCR) may also be used to generate a DNA fragment encoding a protein of interest.

After identifying an appropriate selected gene or DNA molecule, it may be inserted into any one of the many vectors currently known in the art, so that it will direct the expression and production of the selected protein when incorporated into a target cell. In a recombinant expression vector, the coding portion of the DNA segment is positioned under the control of a promoter/enhancer element. The promoter may be in the form of the promoter that is naturally associated with a selected gene, as may be obtained by isolating the 5′ non-coding sequences located upstream of the coding segment or exon, for example, using recombinant cloning and/or PCR technology.

In other embodiments, it is contemplated that certain advantages will be gained by positioning the coding DNA segment under the control of a recombinant, or heterologous, promoter. As used herein, a recombinant or heterologous promoter is intended to refer to a promoter that is not normally associated with a selected gene in its natural environment. Such promoters may include those normally associated with other selected genes, and/or promoters isolated from any other bacterial, viral, eukaryotic, or mammalian cell. Naturally, it will be important to employ a promoter that effectively directs the expression of the DNA segment in the chosen target cells.

The use of recombinant promoters to achieve protein expression is generally known to those of skill in the art of molecular biology, for example, see Sambrook et al (1989; incorporated herein by reference). The promoters employed may be constitutive, or inducible, and can be used under the appropriate conditions to direct high level or regulated expression of the introduced DNA segment. Expression of genes under the control of constitutive promoters does not require the presence of a specific substrate to induce gene expression and will occur under all conditions of cell growth. In contrast, expression of genes controlled by inducible promoters is responsive to the presence or absence of an inducing agent.

Promoters isolated from the genome of viruses that grow in mammalian cells, e.g., RSV, vaccinia virus 7.5K, SV40, HSV, adenoviruses MLP, MMTV LTR and CMV promoters, may be used herewith, as well as promoters produced by recombinant DNA or synthetic techniques. Currently preferred promoters are those such as CMV, RSV LTR, the SV40 promoter alone, and the SV40 promoter in combination with the SV40 enhancer.

Exemplary tissue specific promoter/enhancer elements and transcriptional control regions that exhibit tissue specificity include, but are not limited to: the elastase I gene control region that is active in pancreatic acinar cells; the insulin gene control region that is active in pancreatic cells; the immunoglobulin gene control region that is active in lymphoid cells; the albumin, 1-antitrypsin and -fetoprotein gene control regions that are active in liver; the -globin gene control region that is active in myeloid cells; the myelin basic protein gene control region that is active in oligodendrocyte cells in the brain; the myosin light chain-2 gene control region that is active in skeletal muscle; and the gonadotropic releasing hormone gene control region that is active in the hypothalamus.

Specific initiation signals may also be required for sufficient translation of inserted protein coding sequences. These signals include the ATG initiation codon and adjacent sequences. In cases where the entire coding sequence, including the initiation codon and adjacent sequences are inserted into the appropriate expression vectors, no additional translational control signals may be needed. However, in cases where only a portion of the coding sequence is inserted, exogenous translational control signals, including the ATG initiation codon should be provided. The initiation codon must be in phase with the reading frame of the protein coding sequences to ensure translation of the entire insert. These exogenous translational control signals and initiation codons can be of a variety of origins, both natural and synthetic. The efficiency and control of expression may be enhanced by the inclusion of transcription attenuation sequences, enhancer elements, etc.

A variety of vectors may be used including, but not limited to, those derived from recombinant bacteriophage DNA, plasmid DNA or cosmid DNA. For example, plasmid vectors such as pBR322, pUC 19/18, pUC 118, 119 and the M13 mp series of vectors may be used. Bacteriophage vectors may include gt10, gt11, gt18-23, ZAP/R and the EMBL series of bacteriophage vectors. Cosmid vectors that may be utilized include, but are not limited to, pJB8, pCV 103, pCV 107, pCV 108, pTM, pMCS, pNNL, pHSG274, COS202, COS203, pWE15, pWE16 and the charomid 9 series of vectors. Vectors that allow for the in vitro transcription of RNA, such as SP6 vectors, may also be used to produce large quantities of RNA that may be incorporated into matrices.

The selected genes and DNA segments may also be in the form of a DNA insert located within the genome of a recombinant virus, such as, for example a recombinant herpes virus, retroviruses, vaccinia viruses, adenoviruses, adeno-associated viruses or bovine papilloma virus. While integrating vectors may be used, non-integrating systems, which do not transmit the gene product to daughter cells for many generations will often be preferred. In this way, the gene product is expressed during a defined biological process, e.g., a wound healing process, and as the gene is diluted out in progeny generations, the amount of expressed gene product is diminished.

In such embodiments, to place the gene in contact with a target cell, one would prepare the recombinant viral particles, the genome of which includes the gene insert, and contact the target cells or tissues via release from the polymer fiber of the present invention, whereby the virus infects the cells and transfers the genetic material.

Genes with sequences that vary from those described in the literature are also contemplated for use in the invention, so long as the altered or modified gene still encodes a protein that functions to effect the target cells in the desired (direct or indirect) manner. These sequences include those caused by point mutations, those due to the degeneracies of the genetic code or naturally occurring allelic variants, and further modifications that have been introduced by genetic engineering, i.e., by the hand of man.

Techniques for introducing changes in nucleotide sequences that are designed to alter the functional properties of the encoded proteins or polypeptides are well known in the art. Such modifications include the deletion, insertion or substitution of bases, and thus, changes in the amino acid sequence. Changes may be made to increase the activity of a protein, to increase its biological stability or half-life, to change its glycosylation pattern, confer temperature sensitivity or to alter the expression pattern of the protein, and the like. All such modifications to the nucleotide sequences are encompassed by this invention.

It is an advantage of the present invention that one or more than one selected gene may be used in the gene transfer methods and compositions. The nucleic acid delivery methods may thus entail the administration of one, two, three, or more, selected genes. The maximum number of genes that may be applied is limited only by practical considerations, such as the effort involved in simultaneously preparing a large number of gene constructs or even the possibility of eliciting an adverse cytotoxic effect. The particular combination of genes may be chosen to alter the same, or different, biochemical pathways. For example, a growth factor gene may be combined with a hormone gene; or a first hormone and/or growth factor gene may be combined with a gene encoding a cell surface receptor capable of interacting with the polypeptide product of the first gene.

In using multiple genes, they may be combined on a single genetic construct under control of one or more promoters, or they may be prepared as separate constructs of the same of different types. Thus, an almost endless combination of different genes and genetic constructs may be employed. Certain gene combinations may be designed to, or their use may otherwise result in, achieving synergistic effects on cell stimulation and tissue growth, any and all such combinations are intended to fall within the scope of the present invention. Indeed, many synergistic effects have been described in the scientific literature, so that one of ordinary skill in the art would readily be able to identify likely synergistic gene combinations, or even gene-protein combinations.

It will also be understood that, if desired, the nucleic segment or gene could be administered in combination with further agents, such as, e.g. proteins or polypeptides or various pharmaceutically active agents. So long as genetic material forms part of the composition, there is virtually no limit to other components which may also be included, given that the additional agents do not cause a significant adverse effect upon contact with the target cells or tissues. The nucleic acids may thus be delivered along with various other agents, for example, in certain embodiments one may wish to administer an angiogenic factor as disclosed in U.S. Pat. No. 5,270,300 and incorporated herein by reference.

As the chemical nature of genes, i.e., as a string of nucleotides, is essentially invariant, and as the process of gene transfer and expression are fundamentally the same, it will be understood that the type of genes transferred by the fiber matrices of the present invention is virtually limitless. This extends from the transfer of a mixture of genetic material expressing antigenic or immunogenic fragments for use in DNA vaccination; to the stimulation of cell function, as in wound-healing; to aspects of cell killing, such as in transferring tumor suppressor genes, antisense oncogenes or apoptosis-inducing genes to cancer cells.

By way of example only, genes to be supplied by the invention include, but are not limited to, those encoding and expressing: hormones, growth factors, growth factor receptors, interferons, interleukins, chemokines, cytokines, colony stimulating factors and chemotactic factors; transcription and elongation factors, cell cycle control proteins, including kinases and phosphatases, DNA repair proteins, apoptosis-inducing genes; apoptosis-inhibiting genes, oncogenes, antisense oncogenes, tumor suppressor genes; angiogenic and anti-angiogenic proteins; immune response stimulating and modulating proteins; cell surface receptors, accessory signaling molecules and transport proteins; enzymes; and anti-bacterial and anti-viral proteins.

H. Kits

All the essential materials and reagents required for the various aspects of the present invention may be assembled together in a kit. The kits of the present invention also will typically include a means for containing the vials comprising the desired components in close confinement for commercial sale such as, e.g., injection or blow-molded plastic containers into which the desired vials are retained. Irrespective of the number or type of containers, the kits of the invention are typically packaged with instructions for use of the kit components.
 

Claim 1 of 34 Claims

1. A drug delivery composition comprising at least one fiber having a bore and a wall, wherein said fiber comprises a first component and a second component, and wherein said first component is a biodegradable polymer and said second component is selected from the group consisting of a gel and a hydrogel.
 

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