FIBROTIC TISSUE STABILIZED BY CROSSLINKING FOR TREATMENT OF SNORING AND OBSTRUCTIVE SLEEP APNEA

Abstract

The present invention relates to administration or application of a crosslink inducing agent to a subject following a stiffening procedure to the respiratory system. The present invention has identified that such application or administration of a crosslinking reagent to fibrotic tissue allows for improved stability and strength of the tissue.

Claims

1: A method for improving fibrotic tissue in a subject comprising: a. performing a stiffening procedure on a soft tissue of a subject to induce a fibrotic tissue within the soft tissue; and b. administering in vivo a chemical crosslinking reagent comprising a crosslinker to the fibrotic tissue.

2: The method of claim 1, wherein the chemical crosslinking reagent is administered between 3 days and 3 weeks following the stiffening procedure.

3: The method of claim 2, wherein the chemical crosslinking reagent is administered 3 to 10 days after the stiffening procedure.

4: The method of claim 1, wherein the chemical crosslinking reagent is administered as a solution, spray, gel, encapsulation, or released from a device, coating, patch, strip, suture, implant or combination thereof.

5: The method of claim 1, wherein the chemical crosslinking reagent comprises a crosslinker and a carrier solution.

6: The method of claim 1, wherein the crosslinker is selected from the group consisting of D- or L-Threose, genipin (GP), methylglyoxal (MG), 1-ethyl-3-(3-dimethylamniopropyl) carbodiimide hydrochloride (EDC), proanthrocyanidin, transglutaminase (TG) or combinations thereof.

7: The method of claim 5, wherein the crosslinker is 20-100 mM of D- and/or L-Threose.

8: The method of claim 5, wherein the crosslinker is 5-120 mM of genipin.

9: The method of claim 5, wherein the crosslinker is 5-50 mM of methylglyoxal.

10: The method of claim 5, wherein the crosslinker is 2-50 mM of 1-ethyl-3-(3-dimethylamniopropyl) carbodilimide hydrochloride.

11: The method of claim 5, wherein the crosslinker is 0.025-0.5% w/v of proanthrocyanidin.

12: The method of claim 5, wherein the crosslinker is 0.5-5 U/ml of transglutaminase.

13: The method of claim 1, wherein the carrier solution comprises an additive selected from the group consisting of a buffer, a surfactant, a stabilization agent, a co-factor, a flavoring agent or combinations thereof.

14: The method of claim 1, wherein the chemical crosslinking reagent further comprises a biodegradable or non-degradable polymer to control the release of the crosslinker.

15: The method of claim 1, wherein the stiffening procedure is selected from the group consisting of radio-frequency (RF) heating, radio-frequency ablation (RFA), administration of laser energy, ethanol administration, hypertonic saline administration, sodium tetradecyl sulphate administration, doxycycline administration, insertion of palatal implants or combinations thereof.

16: A method for improving fibrotic tissue in a subject comprising the steps of: preparing a crosslinking reagent by dissolving a crosslinking agent in a carrier medium; and contacting at least a portion of the fibrotic tissue in a subject.

17: The method of claim 16, wherein the contact between the fibrotic tissue and the crosslinking reagent is affected by injections directly into the portion of the fibrotic tissue with a needle.

18: A composition for inducing crosslinking in fibrotic tissue comprising a chemical crosslinking reagent comprising a crosslinker and a carrier solution.

19: The composition of claim 18, wherein the crosslinker comprises 50 mM of genipin.

20: The composition of claim 19, wherein the carrier solution comprises water.

21: The composition of claim 20, wherein the carrier solution further comprises isotonic factors such as phosphate.

22: The method of claim 14, wherein the crosslinker is distributed throughout the biodegradable or non-degradable polymer at a ratio of about 5-100 mg crosslinker to 1 g of polymer.

23: The method of claim 14, wherein the crosslinker comprises genipin and the genipin is distributed throughout the biodegradable or non-degradable polymer at a ratio of about 10 mg genipin to 1 g of polymer.

24: The method of claim 14, wherein the chemical crosslinking reagent is prevented from release from a biodegradable or non-degradable polymer until after an outer coating comprised of a biodegradable polymer has degraded.

Description

DETAILED DESCRIPTION

[0018] The present invention provides for administration or application of a crosslinker to fibrotic tissue. The crosslinker may be administered to fibrotic tissue within a subject through multiple means, either alone or in combination. The crosslinker may be applied or administered as a solution, spray, gel, encapsulation, device, coating, patch, strip, suture, implant or combination thereof.

[0019] Protein crosslinking has been previously used to modify the mechanical properties and chemical stability of collagenous tissues (Charulatha & Rajaram, 2003; Chuang, Odono, & Hedman, 2007; Han, Jaurequi, Tang, & Nimni, 2003; Slusarewicz, Zhu, & Hedman, 2010; Sung, Liang, Chen, Huang, & Liang, 2001; Tang, Sharan, & Vashishth, 2008; Vasudev & Chandy, 1997; Zhai et al., 2006). This has typically been done ex vivo to modulate the strength of tissue implants as well as make them more resistant to enzymatic degradation (Hapach, VanderBurgh, Miller, & Reinhart-King, 2015). Hedman, et al., proposed the use of minimally toxic chemical crosslinkers to treat native tissues in vivo (Chuang, Odono, & Hedman, 2007; Slusarewicz, Zhu, & Hedman, 2010; Slusarewicz, Zhu, Kirking, Toungate, & Hedman, 2011). These prior chemical crosslinking techniques were directed at well-organized but mechanically insufficient or mechanically degraded native tissues, not fibrotic or scar tissues.

[0020] Lau et al. investigated a biological, gene activated matrix approach to generate overexpression of lysyl oxidase to increase crosslinking of collagen and elastin in wound healing applications (Lau, Gobin, & West, 2006). Lysyl oxidase is a native protein that is involved in the reorganization of collagen fibers in granulation tissue by way of inducing directional migration of fibroblasts and catalyzing inter- and intra-molecular covalent crosslinks. In this way the native protein LO is involved in the formation of the fibrotic tissues associated with wound healing and with the remodeling of these tissues. In contrast, a chemical crosslinking approach would act in a non-biologic way to increase the strength and stiffening of the fibrotic or scar tissues and prevent remodeling of these tissues. Essentially chemical crosslinking of fibrotic or scar tissues halts the wound healing process, while augmenting the mechanical properties in such a way as to reduce tissue collapsibility and excessive compliance, whereas LO overexpression promotes the complete wound healing process. In the case of the use of induced fibrosis to stiffen upper airway tissues, the full remodeling process leads eventually to a loss of the mechanical property changes that were the target of the induced fibrosis procedure. And, as has been expressed above, without the timely application of a crosslinking treatment, the beneficial changes brought about by the induced fibrosis are lost to natural remodeling of the fibrotic or scar tissues, even without the overexpression of LO.

[0021] To be an effective in vivo crosslinker, the applied chemical would need to be non-toxic or minimally toxic and react quickly to avoid clearance before action. Such possible chemical crosslinking agents include but are not limited to D- or L-Threose, genipin (GP), methylglyoxal (MG), 1-ethyl-3-(3-dimethylamniopropyl) carbodiimide hydrochloride (EDC), proanthrocyanidin, and transglutaminase (TG). Different crosslinkers have been shown to elicit different mechanical effects on tissues (Slusarewicz et al., 2011) and thus different crosslinkers may be used alone or in combination to modulate the final mechanical properties of the fibrotic tissue or fibrotic and surrounding tissues. Thus the applied solutions, sprays, gels, encapsulations, devices, coatings, patches, strips, sutures, or implants of the present invention may contain at least one chemical crosslinker or a mixture of two or more crosslinking agents. The crosslinkers can be administered in an amount specific to achieve the desired effect and can be administered in a single or multiple administrations. For example, MG can be administered at between about 5 to 50 mM, PA can be administered at between about 0.025 to 0.5% w/v, EDC can be administered at between about 2 to 50 mM, and DT or LT can be administered at between about 20 and 100 mM. Further examples of crosslinkers and their buffers and concentrations can be found in U.S. Pat. No. 8,283,322, which is hereby incorporated by reference in its entirety.

[0022] In the various aspects of the invention, the crosslinker treatment may be applied to a subject at different time points post-surgery, dependent on timeframe for fibrotic tissue formation, preferably within 3 days to 3 weeks but ideally 3-10 days post-surgery.

[0023] In one aspect of the present invention, a solution of crosslinking reagent, in a suitable carrier solution, is injected into the soft palate or tongue base after a stiffening procedure is performed and fibrotic tissue is in place. The nature of methods and devices leading to fibrotic tissue formation can be implant, electrical, light, gas, or chemical based, summarized in Kotecha and Hall (2014) and elsewhere. Chemicals such as sodium tetradecyl sulphate can be injected into the soft palate to create scar tissue and increase, at least temporarily, the stiffness of the tissue which can help alleviate the symptoms of snoring (Brietzke & Mair, 2001). Other agents that can be used to stiffen the soft palate include but are not limited to: ethanol, doxycycline, and hypertonic saline (Brietzke & Mair, 2003, 2004). Radiofrequency ablation (RFA), use of lasers, or RF non-ablative heating are other techniques that involves the creation of scar tissue in the palate region in order to stiffen the palate and reduce snoring (Back, Hytonen, Roine, & Malmivaara, 2009, Neruntarat & Chantapant, 2009). The Pillar procedure involves the implantation of palatal rods in the soft palate with the intention of both augmenting the stiffness of the soft palate by the stiffness of the implants and by the implants and implantation surgery creating scar tissue to stiffen the soft palate (Ho, Wei, & Chung, 2004). As the protein crosslinker reacts with the fibrotic tissue, it can induce crosslink formation that increases the mechanical strength and reduces compliance of the fibrotic and surrounding tissues, and stabilizes the fibrotic or scar tissue and slows resorption and enzymatic degradation. The cros slink augmentation of the present invention can also increase the fibrotic tissue's resistance to subsequent mechanical degradation, which is expected to occur given its poorly organized nature and relatively low strength. The crosslinking reagent can be applied, such by injection, in one location or multiple locations in or around the scar tissue to facilitate a desired distribution of agent or agents and stabilization of fibrotic or scar tissue and surrounding native tissue.

[0024] The carrier solution for the crosslinking reagent can be aqueous or non-aqueous and may contain other non-crosslinking components that may help facilitate crosslinking. Such components include, but are not limited to, buffers (in order to maintain a pH that is optimal to accelerate or extend the crosslinking activity for a particular crosslinker), surfactants (to enhance the distribution of the crosslinker within the tissue), stabilization agents (to maintain the stability of the solution), and co-factors (to enhance the reactivity of the crosslinker). The addition of a flavoring agent may also be used to make the oral treatment more tolerable to patients.

[0025] The crosslinker may be administered to the subject or patient once or over a series of treatments. The crosslinker can be selected from a number of minimally toxic crosslinking agents such as genipin at concentrations between 5-120 mM (preferably about 20-100 mM). Likewise, the buffer can be selected from a number of solutions such as sterile water for injection (with or without a solute to adjust osmolarity), or sterile saline solution, or at a concentration of 25-250 mM 4-(2-Hydroxyethyl)-1-piperazinepropanesulfonic acid (EPPS) (preferably approximately 50 mM) at a pH between 7-10 (preferably about 7.4). The crosslinker reagent can optionally contain 25-250 mM of a phosphate salt (preferably 50 mM) to act as a co-factor and to adjust osmolarity and also optionally contain a co-solvent such as dimethyl sulfoxide (DMSO) in a range between 1-50% (preferably between 10-20%) to increase the solubility of the crosslinking agent.

[0026] In some embodiments, the crosslinker can be methylglyoxal at a concentration ranging from 10-60 mM in sterile water for injection, or in sterile saline solution, or in about 50 mM EPPS buffer, at a pH of about 7.4, and with a solute such as phosphate to adjust osmolarity of the solution.

[0027] In some embodiments, the crosslinker can be proanthrocyanidin at a concentration ranging from 0.025-0.5% w/v in sterile water for injection, or in sterile saline solution, or in about 50 mM EPPS buffer, at a pH of about 7.4, and with a solute such as phosphate to adjust osmolarity of the solution.

[0028] In some embodiments, the crosslinker can be 1-ethyl-3-(3-dimethylamniopropyl) carbodiimide hydrochloride at a concentration ranging from 2-50 mM in about 50 mM 2-(N-morpholino)ethanesulfonic acid (MES) buffer, at a pH of about 6.0.

[0029] In some embodiments, the crosslinker can be L- or D-threose at a concentration ranging from 20-100 mM in sterile water for injection, or in sterile saline solution, or in about 50 mM EPPS buffer, at a pH of about 7.4, and with a solute such as phosphate to adjust osmolarity of the solution.

[0030] In some embodiments, the crosslinker can be transglutaminase at a concentration ranging from 0.5-5 U/ml in sterile water for injection, or in sterile saline solution, or in about 50 mM Tris buffer, at a pH of about 7.4, and with a solute such as phosphate to adjust osmolarity of the solution.

[0031] In another aspect of the present invention, the crosslinking solution described above is administered to the patient in the form of an aerosol spray at crosslinker concentrations and in a buffered carrier similar to that described above. The crosslinking agent can diffuse into the fibrotic tissue in order to stabilize it, improve the mechanical properties, and prevent resorption/degradation. The crosslinker can be delivered from a pump that is pressurized with a suitable gaseous or liquid propellant or one that is actuated using a hand or motorized pump. In this embodiment, the crosslinker could be in the form of lyophilized, sterile tablet or cake that is placed in an aerosol pump containing the carrier solution and shaken until fully solubilized in the solution. One advantage to this delivery method is that it is non-invasive. In addition, the formulation may contain both thickening agents and biocompatible adhesives in order to coat the fibrotic tissue and maintain contact for a prolonged period of time while the crosslinking takes place. The formulation may also contain a surfactant or penetrant to enhance penetration from the surface of the tissue to the underlying tissue. Suitable thickening agents might include, but are not limited to, gellan gum, alginates, agar, carrageenan and pectin, proteins such as gelatin and artificial molecules such as Carbomer (polyacrylic acid), and polyethylene glycol. Non-limiting examples of adhesives might include poly(glycerol-co-sebacate acrylate), oleic methyl esters, or alkyl ester cyanoacrylates. An example of a penetration enhancer includes dimethyl sulfoxide (DMSO).

[0032] In a further aspect of the present invention, the crosslinking agent can be delivered via a patch or a group of patches that is placed on the surface of the fibrotic tissue. The crosslinker can be incorporated into the patch or into a delivery vehicle that coats the surface of the patch that is in contact with the fibrotic tissue. In order to maintain contact with the tissue, the patch may be attached using biocompatible adhesives (such as those described above), biodegradable or semi-permanent sutures, or a tack or staple like device. The patch may contain microneedles for delivery of the crosslinker to the submucosal tissues. The delivery vehicle may be either a suitable solvent, or solution containing appropriate excipients described with or without suitable thickening agents. The formulation may also optionally contain penetration enhancing reagents such as DMSO in order to facilitate the entry of the crosslinker into the tissue. Additionally the formulation can also contain flavoring agents to make the patch more palatable in the patient's mouth. Alternatively the solution may be incorporated into the patch material. This crosslinker may also be in a solid form which would dissolve slowly over time once in contact with bodily fluids, thereby providing a delayed and sustained release of crosslinker over time. In the case of non-degradable polymers, the material of the patch should be porous and the patch would be later removed by a clinician once its function has been completed. In the case of biocompatible non-toxic polymers (such as polylactic or polyglycolic acid), the patch degradation products may be swallowed slowly and cleared by the body as the patch broke down. The crosslinking agent in such an instance should be incorporated into the patch and released to diffuse into the tissue as the patch degrades on the tissue over time. In this embodiment, a patch would not have to be removed by a clinician. In the cases of degradable or non-degradable materials, the patches may be comprised of or coated with a layer or multiple layers of biodegradable polymers that may or may not contain crosslinking agent. Such may facilitate sustained release (for instance in the case of multiple crosslinker containing layers) or delayed release (for instance in the case of a non-crosslinker containing outer layer surrounding a crosslinker containing patch). Different crosslinkers can also be used in different layers of a patch to control the release rates of particular agents over time. These layers may also be used to deliver a flavoring agent to make the patch a more palatable device to be in the mouth while the crosslinking is taking place. In all cases, the patch could be designed to allow airflow in the case of accidental dislodging from the target tissue. This may be achieved using a number of techniques such as; a pattern of perforations and/or intersecting cut-lines that would enable portions of the patch to deform if not attached to the tissue, holes to facilitate airflow, a combination of flaps and holes, by being small enough to not fully obstruct airflow (for instance in the case of a group of smaller patches), or being shaped in such a way that makes blockage unlikely.

[0033] Solid or liquid crosslinkers can be incorporated into a patch by addition of the crosslinker to a molten polymer prior to casting, molding, spinning, or other manufacturing process. Alternatively, the crosslinker can be co-solubilized with the polymer in a suitable solvent (for example, acetone) and then incorporated into the device, such as by solvent evaporation or by precipitation (for example, by the addition of ethanol) of the polymer as described previously (Athanasiou, Singhal, Agrawal, & Boyan, 1995; Singhal, Agrawal, & Athanasiou, 1996). The crosslinker and polymer can also be solubilized separately and mixed prior to precipitation in either the same solvent or different (miscible) solvents. The rate of crosslinker release from the patch can be controlled by varying the concentration of crosslinker as well as by selecting polymer materials with different in vivo degradation profiles.

[0034] The degradation of biodegradable polymers is often associated with a decrease in local pH due to the production of acidic monomers which can negatively affect the efficiency of many protein crosslinkers (Slusarewicz et al., 2010). The acidification of the local region can be counter balanced with the addition of basic salts into the polymer matrix or crosslinker solution/solid (Agrawal & Athanasiou, 1997). Such salts may be inorganic (for example, but not limited to, calcium carbonate, calcium hydroxyapatite, or sodium bicarbonate) or organic (for example, but not limited to, 2-amino-2-hydroxymethyl-propane-,3-diol (Tris) or 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid (HEPES)). These basic salts can be incorporated at a level of about 5-50% by weight of the polymer (preferably 15-30%).

[0035] In another aspect of the present invention, the crosslinker is incorporated into a non-degradable strip-type delivery device, such as those used in a Pillar procedure of the soft palate (Ho, et al. 2004). The crosslinker may be incorporated in any of the ways described herein. The crosslinker may be distributed throughout the non-degradable polymer at a ratio of about 5-100 mg crosslinker to 1 g of polymer. In some embodiments, the crosslinker may be genipin and the genipin may be distributed throughout the non-degradable polymer at a ratio of about 10 mg genipin to 1 g of polymer. Following implantation, the crosslinker can diffuse into and crosslink the fibrous tissue in order to enhance and stabilize the stiffening effects resulting from the induced fibrosis. The crosslinking can help reduce the resorption and mechanical degradation that occurs over time to the unorganized and otherwise mechanically inferior fibrotic or scar tissue.

[0036] In addition, the polymer of the strip may be constructed using a biodegradable polymer, which would allow for a delayed release of the crosslinker after sufficient time for scar formation has been allowed before the strip degrades. The crosslinker can be any of several known minimally toxic crosslinking agents such as genipin or methylglyoxal (preferably about 1-20 mg per strip, although more can be incorporated if needed). The crosslinker may be distributed throughout the biodegradable polymer at a ratio of about 5-100 mg crosslinker to 1 g of polymer. In some embodiments, the crosslinker may be genipin and the genipin may be distributed throughout the biodegradable polymer at a ratio of about 10 mg genipin to 1 g of polymer.

[0037] In another aspect of the invention, the degradable or non-degradable crosslinker delivery device is a suture, or sphere, or pellet, or a number of these devices that is/are inserted into the submucosal region of the tissue. In instances where the device is a stiffening and crosslinker delivery device, the device(s) may be inserted at the time of or after the fibrosis is induced by any method described herein. In the instances that the device is also a means by which the fibrosis is to be generated (as indicated herein), then the time of insertion will determine the onset of fibrosis generation. The delivery device or devices are positioned in the tissue in such a way as to provide crosslinking treatment to the surrounding fibrotic or scar tissue and surrounding intact tissues upon diffusion of the crosslinking agent from the device into the tissue. In another aspect, the device of any type is coated with a crosslinking agent loaded degradable or non-degradable coating and the crosslinker is released over time into the surrounding fibrotic or scar or surrounding intact tissues. In another aspect, the device of any type or geometry which is inserted into the target region of the target tissue is the means by which fibrosis is induced in the targeted tissue, and the same device or a coating on the device contains the crosslinking agent or agents that are released in a delayed fashion so as to form crosslinks in the recently formed fibrotic tissues and surrounding tissues. In another aspect, the delivery device or devices of any type contains the crosslinking agent or agents, and the same device is coated with an outer biodegradable coating that does not contain a crosslinking agent but prevents release of the crosslinker from the underlying device or coating until the outer coating has sufficiently degraded.

[0038] In another aspect of the invention, the polymer coating on strip-type devices used to induce fibrosis would only degrade and release crosslinker after certain environmental conditions are met. This may take the form of a polymer coating that begins degrading around pH 7.4 (normal body pH) and doesn't degrade in the lowered pH of the tissue undergoing wound healing. Alternatively, the crosslinker may only be released from the strip device when certain cytokines are detected or the pH becomes even more acidic with the combination of wound healing and acidic polymeric degradation products (such as lactic and glycolic acid). The biodegradable polymer coating on the strip-type device may also degrade in the presence of end stage wound healing enzymes which function primarily to resorb fibrotic tissue for replacement. Some examples of pH sensitive polymers include, but are not limited to, poly(carboxylic acids), methacrylic acid, methyl methacrylate, and cellulose derived polymers.

[0039] The crosslinker delivery vehicles described herein may be single phase systems consisting of a solution of crosslinking agent either in a solvent or a solvent containing excipients and/or thickening agents. In other aspects of the invention, the carrier vehicle is composed of a two phase system such as an oil in water (O/W) or water in oil (W/O) emulsion. The emulsion may also contain three or more phases such as a water in oil in water (W/O/W) emulsion with the crosslinker incorporated into one or more of the phases as needed. The formulations herein are not to be considered limited to simple emulsion systems and may also include more complex systems such as liposomal formulations and multi-vesicular emulsions.

[0040] In another aspect of the invention, the delivery vehicle is composed of polymeric (degradable or non-degradable) micro or nanospheres which can encapsulate the crosslinking agent. The spheres may serve as a reservoir for the crosslinking agent and release it into the fibrotic tissue as they are degraded or swelled with water.

[0041] Those skilled in the art will appreciate that the claimed invention further comprises compositions and kits for crosslinking fibrotic tissue. Compositions may comprise, for example, a crosslinker as described herein and a carrier solution. The carrier solution may include a buffer, a surfactant, a stabilization agent, a co-factor, a flavoring agent or combinations thereof. The composition may further comprise additives to assist in controlling or delaying the release of the crosslinker, such as biodegradable polymers. Kits may similarly comprise a crosslinker in solution or powder/solid form and optionally a solution to thereafter dissolve in, and a carrier solution. Each component may be separate within the kit, such as in a sterile, isolated environment, such as a vial. The kit may further include directions as to preparing the chemical crosslinking reagent and to how to administer as described herein.

[0042] The present invention may be better understood by referring to the accompanying examples, which are intended for illustration purposes only and should not in any sense be construed as limiting the scope of the invention.

Example 1

[0043] A practitioner can treat a patient who has already had a soft palate stiffening procedure to create fibrotic tissue by a follow-up injection into the soft palate of 50 mM genipin in a buffer solution of sterile water for injection with about 100-120 mM tri-sodium phosphate. The follow-up crosslinker injection can occur 3-10 days following the soft palate stiffening, fibrosis inducing procedure. Sterile water for injection can be injected into a vial containing the sterile genipin and tri-sodium phosphate and then the vial is shaken for 5 minutes or until the solid components are completely solubilized. The injection procedure can be performed with direct observation using smaller volumes injected into multiple sub-mucosal locations in the soft palate.

Example 2

[0044] A practitioner can treat patients who have had a palate stiffening procedure done by applying a crosslinker such as genipin to the fibrotic tissue to stabilize it in the form of an aerosol spray. This crosslinking treatment can be applied 3-10 days following the hard palate stiffening, fibrosis inducing procedure. This genipin spray could be delivered from either a self-containing pressurized container or an actuated pump. The propellant such as a chlorofluorocarbon and the crosslinker can be contained in separate compartments or vessels and the components mixed prior to pressurization or delivery. In the case of an actuated pump, the propellant can be sterile water and the sterile crosslinker in the form of a lyophilized cake can be added to the propellant prior to spraying. The propellant can also contain a thickening agent such as gellan gum, and a biocompatible adhesive such as poly(glycerol-co-sebacate acrylate), and a penetration enhancer such as DMSO.

Example 3

[0045] A practitioner can treat patients who have had a palate stiffening procedure done by applying a patch composed of a non-degradable polymer, polyurethane, that sticks to the fibrotic tissue while releasing crosslinker via microneedles that penetrate the mucosal layer of the soft palate. The patch can be applied 3-10 days following the hard palate stiffening, fibrosis inducing procedure. The patch would need to be removed after a set period of time by either the patient or a trained healthcare professional. The patch is constructed by dispersing solid genipin in the liquid polymer during manufacture at a ratio of 10 mg genipin to 1 g polymer.

Example 4

[0046] A practitioner can treat patients who have had a palate stiffening procedure done by applying a patch composed of a biodegradable polymer comprised of poly (lactic acid) that sticks to the fibrotic tissue while degrading and releasing crosslinker. The patch can be applied 3-10 days following the hard palate stiffening, fibrosis inducing procedure. No removal of the patch would be necessary as the material would slowly degrade as it released crosslinker and the degradation products would be swallowed and cleared by the patient's body. The patch is constructed by dispersing solid genipin in the liquid polymer during manufacture at a ratio of 10 mg genipin to 1 g polymer. The tissue contacting surface of the patch can contain a biocompatible adhesive such as poly(glycerol-co-sebacate acrylate).

Example 5

[0047] A practitioner can treat patients who plan to have a palate stiffening procedure done, similar to the Pillar Procedure, where non-degradable polymer strips are implanted to induce scar tissue formation. These non-degradable polymer strips can be coated in a degradable polymer, such as poly(lactic or poly(glycolic acid), that contains approximately 10 mg of genipin to be released into the tissue as the polymer degrades. The polymer strips can be coated by either immersing them in molten coating material containing the crosslinking agent or by repeated immersions in a polymer-solvent-crosslinker solution in which the solvent could be evaporated off leaving only the coating material and crosslinker on the strips.

Example 6

[0048] A practitioner can treat patients who have had a palate stiffening procedure done by inserting genipin coated suture(s) into the fibrotic tissue in order to crosslink it. The sutures with genipin loaded coatings can be inserted 3-10 days following the hard palate stiffening, fibrosis inducing procedure. The coating can be put on the suture by immersing the suture in a polymer (poly(lactic acid or poly(glycolic acid)) that has been solubilized in a solvent along with the crosslinking agent genipin at a concentration around 50 mM. The solvent can be evaporated off leaving only the coating and genipin on the suture. These sutures could be degradable like the coating such that no second procedure is required to retrieve the delivery vehicle (suture/suture coating).

Example 7

[0049] A practitioner can treat patients by inserting genipin coated suture(s) into the fibrotic tissue in order to both induce a fibrotic response and to crosslink the fibrotic tissue. The coating can be put on the suture by immersing the suture in a polymer (poly(lactic acid or poly(glycolic acid)) that has been solubilized in a solvent along with the crosslinking agent genipin at a concentration around 50 mM. The solvent can be evaporated off leaving only the coating and genipin on the suture. These sutures could be degradable like the coating such that no second procedure is required to retrieve the delivery vehicle (suture/suture coating).

Example 8

[0050] A practitioner can treat patients who have had a tongue base stiffening procedure done by injecting or spraying genipin at a concentration around 50 mM in order to stabilize the fibrotic tissue and retain its beneficial effects for a longer period of time. The injection or spraying can be performed 3-10 days following the hard palate stiffening, fibrosis inducing procedure. The addition of about 50-100 mM phosphate solution can be added to the genipin in the case of an injection in order to increase the amount of crosslinking. In the case of an aerosol spray, a propellant such as a chlorofluorocarbon can be included with or without 50-100 mM phosphate to facilitate delivery and crosslinking in the target fibrotic tissue.

[0051] The foregoing examples have been presented for purposes of illustration and description. They are not intended to be exhaustive or to limit the embodiments to the precise forms disclosed. Obvious modifications and variations are possible in light of the above teachings. All such modifications and variations are within the scope of the appended claims when interpreted in accordance with the breadth to which they are fairly, legally and equitably entitled. All references to patent related and non-patent related literature are hereby incorporated by reference in their entirety.

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