Maximizing distribution and minimizing washout of injectates in bone and method for enhanced aspirate extraction

Abstract

To distribute agents into bone and maintain those agents within bone.SOLUTION: These methods include mechanical, rheological, pharmacological, and other methods. By maintaining injectates within a targeted location in bone and increasing their distribution in bone, the methods are useful for purposes including but not limited to increasing local concentration, improving therapeutic effectiveness, increasing duration of action, and decreasing systemic toxicity. These same methods can displace the contents of bone toward a harvesting instrument for purposes of collection with similar and complimentary methods. The methods are useful for controlling the rate of egress of intraosseous distribution of agents or cells for a variety of purposes, including but not limited to: augmenting stem cell recovery in bone marrow aspiration; chemotherapy; medication delivery; treatment of infection; bone augmentation; and the like.SELECTED DRAWING: None

Description

[Technical Field]

[0001] (CROSS-REFERENCE TO RELATED APPLICATIONS) This application claims priority to U.S. Provisional Patent Application No. 62 / 844,592, filed May 7, 2019, the contents of which are incorporated by reference herein in their entirety. [Background technology]

[0002] Intraosseous (IO) and infusion administration are primarily used in emergency situations when intravenous administration is difficult or impossible. Because bone veins are fragile with imperfect walls and bone is a rigid body, drugs administered intraosseously under high pressure rapidly reach the bone and enter the systemic circulation in a manner similar to intravenous (IV) administration when flushed with saline. Indeed, for nearly all purposes, IO administration is considered equivalent to IV administration. For example, the first study of IO administration used iodinated contrast in dogs and imaged them with fluoroscopy, observing that the contrast reached the heart in 4 seconds. Specific pharmacological studies have been conducted with certain drugs that support this assumption. The IO route is typically used for resuscitation, and vasopressors are used in advanced cardiopulmonary resuscitation (ACLS) and pediatric advanced life support (PALS) resuscitation.

[0003] However, immediate systemic distribution of intraosseously administered drugs is not required in all procedures. Bone-targeted therapy loses effectiveness if the therapeutic agent cannot localize to the target bone. Sample collection may be diluted or reduced if the desired portion is shed from the bone. In some cases, treatment may be dangerous if the administered drug is present in the bone and may lead to systemic toxicity. Examples include, but are not limited to, stem cell collection, intraosseous infection, tumor treatment, and bone stabilization (e.g., vertebroplasty).

[0004] Therefore, there is a need in the art for improved methods of maintaining implants within bone, and the present invention meets this need. Summary of the Invention

[0005] In one aspect, the present invention relates to a method for controlling the release rate and intraosseous distribution of an injectate, comprising the steps of providing a quantity of at least one injectate comprising at least one therapeutic agent; injecting the quantity of the at least one injectate into a first bone portion via a first lumen; and maintaining the quantity of the at least one injectate in a localized state within the first bone portion such that at least 15% to 95% of the quantity is retained for at least 5 minutes to 2 hours.

[0006] In one embodiment, the at least one injectate further comprises at least one vasoactive agent. In one embodiment, the injecting step is performed after the step of administering at least one first injectate at a pressure or rate such that connections between the peripheral veins of the first bone portion are opened through incomplete or fragile sinusoidal walls. In one embodiment, the pressure is selected from the group consisting of about 10 psi, 20 psi, 30 psi, 40 psi, 50 psi, 60 psi, 70 psi, 80 psi, 90 psi, 100 psi, 200 psi, 300 psi, 400 psi, 500 psi, 600 psi, 700 psi, 800 psi, 900 psi, and 1000 psi. In one embodiment, the rate is selected from the group consisting of about 0.1 μL / s, 1 μL / s, 10 μL / s, 50 μL / s, 0.1 mL / s, 0.2 mL / s, 0.3 mL / s, 0.4 mL / s, 0.5 mL / s, 0.6 mL / s, 0.7 mL / s, 0.8 mL / s, 0.9 mL / s, 1 mL / s, 2 mL / s, 3 mL / s, 4 mL / s, 5 mL / s, 6 mL / s, 7 mL / s, 8 mL / s, 9 mL / s, and 10 mL / s.

[0007] In one embodiment, the amount of at least one vasoactive agent is injected before the amount of the at least one infusate. In one embodiment, the amount of at least one vasoactive agent is injected after the amount of the at least one infusate. In one embodiment, the amount of at least one vasoactive agent is injected simultaneously with the amount of the at least one infusate. In one embodiment, the amount of at least one vasoactive agent is injected adjacent to a draining vein of the bone. In one embodiment, the first bone portion is located non-adjacent to a draining vein of the bone.

[0008] In one embodiment, the at least one vasoactive agent is a vasoconstrictor selected from the group consisting of a sympathomimetic agent, methoxamine hydrochloride, epinephrine, dobutamine, dopamine, norepinephrine, milrinone, midodrine hydrochloride, desglimidodrine, and an alpha receptor agonist, stimulator, or activator. In one embodiment, the at least one vasoactive agent is a vasodilator selected from the group consisting of an α- or β-adrenergic receptor modulator, a muscarinic agent, a histamine agent, a dopamine receptor modulator, an organic nitrate, isosorbide mononitrate, a mononitrate, isosorbide dinitrate, a dinitrate, nitroglycerin, a trinitrate, minoxidil, sodium nitroprusside, hydralazine hydrochloride, nitric oxide, nicardipine hydrochloride, fenoldopam mesylate, diazoxide, enalaprilat, epoprostenol sodium, a prostaglandin, milrinone lactate, a bipyridine, and a dopamine D1-like receptor agonist, stimulator, or activator. In one embodiment, the at least one therapeutic agent is selected from the group consisting of an anti-tumor agent, an anti-proliferative agent, an angiogenesis inhibitor, a cytotoxic agent, an anti-microbial agent (including an anti-bacterial agent and an anti-fungal agent), a cell mobilizer, and an analgesic.

[0009] In one embodiment, the method further comprises the step of selecting an infusion rate of the at least one infusate from the group consisting of about 0.1 μL / s, 1 μL / s, 10 μL / s, 50 μL / s, 0.1 mL / s, 0.2 mL / s, 0.3 mL / s, 0.4 mL / s, 0.5 mL / s, 0.6 mL / s, 0.7 mL / s, 0.8 mL / s, 0.9 mL / s, 1 mL / s, 2 mL / s, 3 mL / s, 4 mL / s, 5 mL / s, 6 mL / s, 7 mL / s, 8 mL / s, 9 mL / s, and 10 mL / s. In one embodiment, the volume of the at least one infusate is selected from the group consisting of about 0.1 μL, 1 μL, 10 μL, 50 μL, 100 μL, 500 μL, 1 mL, 2 mL, 3 mL, 4 mL, 5 mL, and 10 mL. In one embodiment, the amount of the at least one vasoactive agent is selected to reduce blood flow at the bone by about 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, or 95%. In one embodiment, the amount of at least one therapeutic agent has a concentration selected from the group consisting of greater than about 0% by weight, greater than about 5% by weight, greater than about 10% by weight, greater than about 15% by weight, greater than about 20% by weight, greater than about 25% by weight, greater than about 30% by weight, greater than about 35% by weight, greater than about 40% by weight, greater than about 45% by weight, greater than about 50% by weight, greater than about 55% by weight, greater than about 60% by weight, greater than about 65% by weight, greater than about 70% by weight, greater than about 75% by weight, greater than about 80% by weight, greater than about 85% by weight, greater than about 90% by weight, or greater than about 95% by weight.

[0010] In one aspect, the method further includes, after the step of maintaining the at least one injectate volume within the first bone portion in a localized state, extracting an aspirate volume from one or more additional bone portions via one or more additional lumens, such that a pressure gradient is formed across the vasculature or bone region between the first bone portion having a first pressure and the one or more additional bone portions, each having a pressure lower than the first pressure.

[0011] In one embodiment, the first lumen and the one or more additional lumens are disposed within a single device. In one embodiment, the first lumen is disposed within a first device, and the one or more additional lumens are disposed within one or more additional devices. In one embodiment, the one or more additional bone segments are each spaced from the first bone segment by about 5 mm to about 500 mm. In one embodiment, the injecting procedure and the extracting procedure are performed at sequential depths in their corresponding bone segments.

[0012] In another aspect, the present invention relates to a method for increasing aspirant yield from a bone portion, comprising the steps of providing at least one injectant amount; injecting the at least one injectant amount into a first bone portion via a first lumen; and extracting an aspirant amount from one or more additional bone portions via one or more additional lumens; wherein the first bone portion and the second bone portion are located between about 5 and 500 mm from each other; and a pressure gradient is formed across a region of vasculature or bone between the first bone portion having a first pressure and the one or more additional bone portions each having a pressure lower than the first pressure.

[0013] In one embodiment, the injecting and extracting procedures are performed simultaneously. In one embodiment, the extracting procedure is performed consecutively after the injecting procedure. In one embodiment, the injecting and extracting procedures are performed at sequential depths in their corresponding bone portions. In one embodiment, the first lumen and the one or more additional lumens are disposed within a single device. In one embodiment, the first lumen is disposed within a first device and the one or more additional lumens are disposed within one or more additional devices.

[0014] In one embodiment, the method further comprises the step of selecting the positive infusion pressure and the negative extraction pressure from the group consisting of about 10 psi, 20 psi, 30 psi, 40 psi, 50 psi, 60 psi, 70 psi, 80 psi, 90 psi, 100 psi, 200 psi, 300 psi, 400 psi, 500 psi, 600 psi, 700 psi, 800 psi, 900 psi, and 1000 psi. In one embodiment, the method further comprises the step of selecting the infusion and extraction rates from the group consisting of about 0.1 μL / s, 1 μL / s, 10 μL / s, 50 μL / s, 0.1 mL / s, 0.2 mL / s, 0.3 mL / s, 0.4 mL / s, 0.5 mL / s, 0.6 mL / s, 0.7 mL / s, 0.8 mL / s, 0.9 mL / s, 1 mL / s, 2 mL / s, 3 mL / s, 4 mL / s, 5 mL / s, 6 mL / s, 7 mL / s, 8 mL / s, 9 mL / s, and 10 mL / s.

[0015] In one embodiment, the at least one injectate volume and the aspirate volume are each selected from the group consisting of about 0.1 μL, 1 μL, 10 μL, 50 μL, 100 μL, 500 μL, 1 mL, 2 mL, 3 mL, 4 mL, 5 mL, and 10 mL. In one embodiment, the aspirate volume is a percentage of the at least one injectate volume, the percentage being selected from the group consisting of 50%, 60%, 70%, 80%, 90%, and 100%. In one embodiment, the at least one injectate volume comprises a fluid. In one embodiment, the at least one injectate volume comprises at least one fluid agent selected from a vasoactive agent, an anti-tumor agent, an anti-proliferative agent, an angiogenesis inhibitor, a cytotoxic agent, an anti-microbial agent (including anti-bacterial and anti-fungal agents), a cell mobilizing agent, and an analgesic agent.

[0016] In one embodiment, the vasoactive agent is a vasoconstrictor selected from the group consisting of a sympathomimetic agent, methoxamine hydrochloride, epinephrine, dobutamine, dopamine, norepinephrine, milrinone, midodrine hydrochloride, desglimidodrine, and an alpha receptor agonist, stimulator, or activator. In one embodiment, the vasoactive agent is a vasodilator selected from the group consisting of alpha or beta adrenergic receptor modulators, muscarine, histamine, dopamine receptor modulators, organic nitrates, isosorbide mononitrate, mononitrates, isosorbide dinitrate, dinitrates, nitroglycerin, trinitrates, minoxidil, sodium nitroprusside, hydralazine hydrochloride, nitric oxide, nicardipine hydrochloride, fenoldopam mesylate, diazoxide, enalaprilat, epoprostenol sodium, prostaglandins, milrinone lactate, bipyridine, and dopamine D1-like receptor agonists, stimulators, or activators. In one embodiment, the at least one fluid agent has a concentration selected from the group consisting of greater than about 0% by weight, greater than about 5% by weight, greater than about 10% by weight, greater than about 15% by weight, greater than about 20% by weight, greater than about 25% by weight, greater than about 30% by weight, greater than about 35% by weight, greater than about 40% by weight, greater than about 45% by weight, greater than about 50% by weight, greater than about 55% by weight, greater than about 60% by weight, greater than about 65% by weight, greater than about 70% by weight, greater than about 75% by weight, greater than about 80% by weight, greater than about 85% by weight, greater than about 90% by weight, or greater than about 95% by weight.

[0017] In another aspect, the present invention relates to a method for localizing an injectate within a bone portion, comprising the steps of: providing at least one double-lumen needle having a first lumen terminating in a distal opening and a second lumen having at least one side port proximate the opening of the first lumen; inserting the at least one double-lumen needle into a target bone portion having a dosing area and a vasoconstrictor area such that the distal opening of the first lumen is positioned toward the vasoconstrictor area and the at least one side port of the second lumen is positioned toward the dosing area; administering a vasoconstrictor through the first lumen and the opening such that the vasoconstrictor localizes within the vasoconstrictor area and reduces blood, cell, or injectate shedding from the dosing area; and administering one or more injectates through the second lumen and the at least one side opening such that the one or more injectates are localized in the dosing area.

[0018] In one aspect, the method further comprises extracting tissue using the at least one double-lumen needle through the at least one side port of the second lumen. In one aspect, at least one additional needle is provided, the needle having at least one lumen. In one aspect, the at least one additional needle is a single-lumen needle. In one aspect, the method further comprises inserting the at least one additional needle into at least one bone portion proximate the target bone portion and aspirating using the at least one additional needle. In one aspect, the at least one additional needle is a double-lumen needle having a first lumen terminating in a distal opening and a second lumen with at least one side port proximate the opening of the first lumen. In one aspect, the method further comprises inserting the at least one additional double-lumen needle into at least one bone portion proximate the target bone portion and administering a vasoconstrictor using the first lumen and the distal opening of the at least one additional double-lumen needle. In one aspect, the method further includes inserting the at least one additional double-lumen needle into at least one bone portion adjacent to the target bone portion and aspirating using the lumen of the at least one additional double-lumen needle. [Brief explanation of the drawings]

[0019] The following detailed description of embodiments of the present invention will be better understood when read in conjunction with the accompanying drawings, it being understood, however, that the invention is not limited to the precise arrangements and instrumentalities of the embodiments shown in the drawings.

[0020] [Figure 1] 10 is a flow chart illustrating an exemplary method of maintaining an injectate within a bone.

[0021] [Figure 2] 10 is a flow chart illustrating another exemplary method of maintaining an injectate within a bone.

[0022] [Figure 3] 10 is a flow chart illustrating an exemplary method of localizing an injectate into a bone.

[0023] [Figure 4] A schematic diagram of an exemplary double lumen needle (left) and a close-up of the needle tip of the double lumen needle (right) showing the distal opening to the first lumen and several closable openings to the second lumen by varying their positions are shown.

[0024] [Figure 5] 10 shows a schematic representation of preparing a target bone for sustaining an injectate into the bone using a double lumen needle.

[0025] [Figure 6] 1 shows a diagram illustrating an exemplary method of maintaining an injectate within a bone.

[0026] [Figure 7] Fluoroscopic image of contrast agent administered into the bone of a subject at a maximum distribution rate of 0.5 mL / sec. The administration rate combined with the viscosity shown does not adequately displace the flowing blood volume within the bone sinusoids.

[0027] [Figure 8] Fluoroscopic image of contrast agent administered into the bone of a subject at a maximum distribution rate of 10 mL / sec. The administration rate is so high that the contrast agent immediately enters the bloodstream without being retained within the bone.

[0028] [Figure 9] Fluoroscopic image of contrast agent co-administered with epinephrine into the subject's bone at a maximum distribution rate of 5 mL / sec. Note the constricted intraosseous draining venules and veins. The placement of the administration needle close to the draining central vein reveals a relatively small bone perfusion area.

[0029] [Figure 10]Fluoroscopic image of contrast agent administered without epinephrine into the bone of a subject at a maximum distribution rate of 5 mL / sec. The distance of the injection site from the draining central vein is maximized. Note the larger area of ​​drug opacification.

[0030] [Figure 11] Fluoroscopic image of contrast agent administered with epinephrine into the bone of a subject at a maximum distribution rate of 5 mL / sec. The distance of the injection site from the draining central vein is maximized. Although the clearance rate of the contrast agent is reduced, the area of ​​clearing is not as large, likely due to the constricted veins.

[0031] [Figure 12] Fluoroscopic image 20 minutes after injection of contrast agent administered with epinephrine into the bone of a subject at a maximum distribution rate of 5 mL / sec. The contrast agent containing epinephrine remains in the bone for 20 minutes, despite a smaller perfusion area. DETAILED DESCRIPTION OF THE INVENTION

[0032] The present invention provides for the distribution and retention of drugs within bone. These methods include mechanical, rheological, pharmacological, and other methods. By maintaining injectates within targeted sites within bone and increasing their distribution within bone, the methods are useful for purposes including, but not limited to, increasing local concentrations, improving therapeutic efficacy, increasing duration of action, and reducing systemic toxicity. These same methods can also transfer intraosseous contents to a collection device for collection purposes by similar and complementary methods. In some embodiments, the methods are useful for setting and distributing structural augmentation materials, such as cements or biologics, to augment bone. In some embodiments, the methods are useful for reducing cell release from the medullary cavity. In some embodiments, the methods are useful for transferring bone marrow contents to a collection device. In some embodiments, the methods are useful for treating vascular or neurological diseases within bone, such as certain types of arthritis. In some embodiments, the methods are useful for treating bone disorders associated with abnormal intraosseous pressure. Without the risk of systemic distribution, the method allows for greater amounts than would be systemically tolerated to be administered intraosseously, retained within the bone, and optionally aspirated or effluxed with one or more devices. The method is useful for controlling the release rate of intraosseous distribution of drugs or cells for a variety of purposes, including, but not limited to, increasing stem cell recovery in bone marrow aspirates, chemotherapy, drug delivery, treatment of infection, bone augmentation, etc.

[0033] definition

[0034] It should be understood that the figures and descriptions of the present invention are simplified to illustrate elements relevant to a clear understanding of the invention, while excluding, for purposes of clarity, many other elements typically found in the art. One of ordinary skill in the art may recognize that other elements and / or procedures are desirable and / or required in practicing the present invention. However, because such elements and procedures are well known in the art, and because they do not facilitate a better understanding of the invention, a discussion of such elements and procedures is not provided herein. The disclosure herein is directed to all such variations and modifications to such elements and methods known to those of ordinary skill in the art.

[0035] Unless defined elsewhere, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. Any methods and materials similar or equivalent to those described herein can be used in the practice or testing of the present invention, and exemplary methods and materials are described.

[0036] As used herein, each of the following terms has the meaning associated with it in this section.

[0037] The articles "a" and "an" are used herein to refer to one or to more than one (i.e., to at least one) of the grammatical object of the article. For example, "an element" means one element or more than one element.

[0038] As used herein, "about" when referring to a measurable value such as an amount, a time period, etc., is meant to include variations of ±20%, ±10%, ±5%, ±1%, and ±0.1% from the stated value, as such variations are appropriate.

[0039] As used herein, the term "anti-tumor effect" refers to a biological effect that may be manifested by a reduction in tumor volume, a reduction in the number of tumor cells, a reduction in the number of metastases, an increase in life expectancy, or an amelioration of various physiological symptoms associated with a cancerous condition. An "anti-tumor effect" may also be manifested by the ability of the compositions of the present invention to prevent the development of a tumor at the primary site.

[0040] A "disease" is a state of health in an animal in which homeostasis cannot be maintained, and where, if the disease is not reversed, the animal's health continues to deteriorate. In contrast, an animal's "disorder" is a state of health in which the animal can maintain homeostasis, but the animal's health is less favorable than it would be without the disorder. If left untreated, the disorder does not necessarily further deteriorate the animal's health.

[0041] As used herein, the term "inhibit" means to reduce or block an activity or function by at least about 10 percent compared to a control value. For example, the activity may be reduced or blocked by 50%, 70%, or 95% compared to a control value.

[0042] The terms "treatment," "treating," and the like, are used herein to generally mean obtaining a desired pharmacological and / or physiological effect. The effect may be prophylactic, in that it completely or partially prevents the disease or its symptoms, and / or therapeutic, in that it partially or completely treats the disease and / or adverse effects caused by the disease. As used herein, the term "treatment" extends to any treatment of a subject's disease and includes: (a) preventing a disease associated with a desired immune response from occurring in a subject who may be predisposed to the disease; (b) inhibiting the disease, i.e., halting its development; or (c) alleviating the disease, i.e., causing the disease to regress.

[0043] "Effective amount" and "pharmaceutically effective amount" refer to a sufficient amount of a drug to provide a desired biological result, which may be a reduction and / or alleviation of the signs, symptoms, or causes of a disease or disorder, or any other desired alteration of a biological system. An appropriate effective amount in any individual case may be determined by one of ordinary skill in the art using routine experimentation.

[0044] "Therapeutically effective amount" refers to that amount that provides a therapeutic effect for the indicated condition and administration regimen. In particular, a "therapeutically effective amount" refers to an amount that prevents, alleviates, or ameliorates symptoms of the disease or prolongs the survival of the treated subject, which may be a human or non-human animal. Determination of a therapeutically effective amount is within the skill of one of ordinary skill in the art.

[0045] As used herein, the term "pharmaceutical composition" refers to a mixture of at least one compound of the present invention with other chemical components and entities, such as carriers, stabilizers, diluents, dispersing agents, suspending agents, thickeners, and / or excipients. The pharmaceutical composition facilitates administration of the compound to an organism. Numerous techniques for administering compounds exist in the art, including, but not limited to, intravenous, oral, aerosol, parenteral, ocular, pulmonary, and topical administration.

[0046] "Pharmaceutically acceptable" means those properties and / or substances that are acceptable to a patient from a pharmacological / toxicological standpoint with respect to composition, formulation, stability, patient acceptability and bioavailability, and to a pharmaceutical chemist in the manufacturing industry from a physical / chemical standpoint. A "pharmaceutically acceptable carrier" means a medium that does not interfere with the effectiveness of the biological activity of the active ingredient and that is not toxic to the host to which it is administered.

[0047] As used herein, "pharmaceutically acceptable carrier" means a pharmaceutically acceptable material, composition, or carrier, such as a liquid or solid filler, stabilizer, dispersing agent, suspending agent, diluent, excipient, thickener, solvent, or encapsulating material, involved in carrying or transporting a compound useful in the present invention in or to a patient so that it may perform its intended function. Typically, such compositions are carried or transported from one organ or part of the body to another. Such a carrier must be "acceptable" in the sense of being compatible with the other ingredients of the formulation, including the compound useful in the present invention, and not harmful to the patient. Some examples of materials which can serve as pharmaceutically acceptable carriers include sugars such as lactose, glucose, and sucrose, starches such as corn starch and potato starch; cellulose and its derivatives, such as sodium carboxymethylcellulose, ethyl cellulose, and cellulose acetate; powdered tragacanth, malt; gelatin; talc; excipients such as cocoa butter and suppository waxes; oils such as peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil, corn oil, and soybean oil; glycols such as polyethylene glycol; polyols such as glycerin, sorbitol, mannitol, and polyethylene glycol; esters such as ethyl oleate and ethyl laurate; agar; buffers such as magnesium hydroxide and aluminum hydroxide; surfactants; alginic acid; pyrogen-free water; isotonic saline; Ringer's solution; ethyl alcohol; phosphate buffers; and other non-toxic, compatible substances used in pharmaceutical formulations. Also, as used herein, "pharmaceutically acceptable carrier" includes any and all coatings, antimicrobials (e.g., antibacterial and antifungal agents), absorption delaying agents, and the like that are compatible with the activity of the compounds useful in the present invention and are physiologically acceptable to the patient. Additional active compounds may also be incorporated into the compositions. "Pharmaceutically acceptable carrier" also includes pharmaceutically acceptable salts of the compounds useful in the present invention.

[0048] The terms "patient," "subject," "individual," and the like are used interchangeably herein and refer to any animal, or cells thereof, either in vitro or in situ, according to the methods described herein. In one non-limiting embodiment, the patient, subject, or individual is a human.

[0049] The phrase "biological sample" is used herein in its broadest sense. A sample can be any biological tissue or fluid in which a biomarker of the present invention can be detected, extracted, isolated, characterized, or measured. Examples of such samples include, but are not limited to, blood, lymph, urine, gynecological fluids, biopsies, amniotic fluid, and smears. Samples that are liquid in nature are referred to herein as "body fluids." Biological samples can be obtained from a patient by various techniques, such as by scraping or swabbing an area or by using a needle to aspirate a body fluid. Methods for collecting various biological samples are well known in the art. Often, the sample will be a "clinical sample," i.e., a sample derived from a patient. Such samples include, but are not limited to, body fluids that may or may not contain cells, such as blood (e.g., whole blood, serum, or plasma), urine, saliva, tissue or fine needle biopsy samples, and archival samples with known diagnostic, treatment, and / or outcome histories. Biological samples also include tissues, such as frozen sections obtained for histological purposes. They also include any material derived from a biological sample by processing the sample. Derived materials include, but are not limited to, cells (or their progeny) isolated from the sample, proteins, or nucleic acid molecules extracted from the sample. Processing a biological sample can include one or more of filtration, distillation, extraction, concentration, inactivation of interfering components, addition of reagents, etc.

[0050] As used herein, the term "injectate" refers to any injectable material that is typically capable of traversing an injection site through a delivery lumen and is generally provided as a gas, liquid, solution, gel, mixture, suspension, cement, or viscous paste.

[0051] Throughout this disclosure, various aspects of the invention may be presented in a range format. It should be understood that descriptions in range format are merely for convenience and brevity and should not be construed as an inflexible limitation on the scope of the invention. Accordingly, descriptions of ranges should be construed as including all possible subranges and individual numerical values ​​within those ranges that are specifically disclosed. For example, a description of a range such as 1 to 6 should be construed as specifically disclosed subranges such as 1 to 3, 1 to 4, 1 to 5, 2 to 4, 2 to 6, 3 to 6, etc., and individual numbers within that range, e.g., 1, 2, 2.7, 3, 4, 5, 5.3, 6, and any whole or partial increments therebetween.

[0052] 1, an exemplary method 100 is described for controlling the release rate and intraosseous distribution of an injectate. Method 100 begins with step 102, in which a quantity of at least one injectate including at least one therapeutic agent is provided. In step 104, the quantity of the at least one injectate is injected into a first bone portion via a first lumen. In step 106, the at least one injectate is maintained within the first bone portion in a localized state.

[0053] In some embodiments, the steps are performed in the listed order. In some embodiments, the steps are performed after an optional first step of administering at least one first injectate at a pressure or rate such that connections between the peripheral veins of the first bone portion are opened through incomplete or fragile sinusoidal walls. The injectate can be any of the drugs described elsewhere herein. Non-limiting examples of injectates include normal saline (about 0.9%) or diluted heparinized solutions. The administration pressure can be about 10 psi, 20 psi, 30 psi, 40 psi, 50 psi, 60 psi, 70 psi, 80 psi, 90 psi, 100 psi, 200 psi, 300 psi, 400 psi, 500 psi, 600 psi, 700 psi, 800 psi, 900 psi, and 1000 psi. The administration rate can be about 0.1 μL / s, 1 μL / s, 10 μL / s, 50 μL / s, 0.1 mL / s, 0.2 mL / s, 0.3 mL / s, 0.4 mL / s, 0.5 mL / s, 0.6 mL / s, 0.7 mL / s, 0.8 mL / s, 0.9 mL / s, 1 mL / s, 2 mL / s, 3 mL / s, 4 mL / s, 5 mL / s, 6 mL / s, 7 mL / s, 8 mL / s, 9 mL / s, and 10 mL / s.

[0054] Also, in some embodiments, an amount of at least one vasoactive agent is provided. The at least one vasoactive agent and the at least one infusate, alone or in combination, may be provided with or without systemic delivery of a therapeutic substance that may further enhance the function of the intraosseously administered drug. In some embodiments, the at least one vasoactive agent is injected before injecting the amount of the at least one infusate. In some embodiments, the at least one vasoactive agent is injected after injecting the amount of the at least one infusate. In some embodiments, the vasoactive agent is injected simultaneously with the amount of the at least one infusate. The sites of injection of the at least one vasoactive agent and the at least one infusate can be the same site or different sites. Possible sites include near a draining vein of the bone or non-adjacent to a draining vein of the bone. Vasoactive agent compositions, therapeutic drug compositions, injection rates, injection volumes, injection concentrations, and injection sites are described in detail elsewhere herein.

[0055] Therapeutic drugs

[0056] The present invention encompasses the use of any suitable therapeutic agent. Some therapeutic agents may be more suitable than others depending on the procedure and the desired effect. In various embodiments, the therapeutic agent is useful for treating a disease or disorder. In various embodiments, the therapeutic agent is useful for obtaining tissue or cell samples.

[0057] In some embodiments, the therapeutic agent comprises a biologic, e.g., a solution or matrix containing biologically active cells, which may be autologous or allogeneic cells, either of which may be cultured, expanded, differentiated, or genetically altered in any manner. In some embodiments, the therapeutic agent comprises a solution or matrix that can attract cells via cell signaling molecules.

[0058] In some embodiments, the therapeutic drug is useful for treating cancer. Therapeutic agents for treating cancer include antitumor and chemotherapeutic agents, such as cytotoxic agents (e.g., 5-fluorouracil, cisplatin, carboplatin, methotrexate, daunorubicin, doxorubicin, vincristine, vinblastine, oxorubicin, carmustine (BCNU), lomustine (CCNU), cytarabine USP, cyclophosphamide, estramucine phosphate sodium, altretamine, hydroxyurea, ifosfamide, procarbazine, mitomycin, busulfan, cyclophosphamide, mitoxantrone, carboplatin, cisplatin, interferon alpha 2A recombinant, paclitaxel, teniposide, and streptozocin), cytotoxic alkylating agents (e.g., busulfan, chlorambucil, cyclophosphamide, melphalan, or ethylesulfonic acid). acid), alkylating agents (e.g., asaley, AZQ, BCNU, busulfan, bisulfan, carboxyphthalatoplatinum, CBDCA, CCNU, CHIP, chlorambucil, chlorozotocin, cis-platinum, clomesone, cyanomorpholinodoxorubicin, cyclodisone, cyclophosphamide, dianhydrogalactitol, fluorodopan, hepsulfam, hycanthone, ifosfamide, melphalan, methyl CCNU, mitomycin C, mitozolamide, nitrogen mustard, PCNU, piperazine, piperazinedione, pipobroman, porfiromycin, spirohydantoin mustard, streptozotocin, teroxylon, tetraplatin, thiotepa, triethylenemelamine, uracil nitrogen mustard, and Yoshi-864), mitotic inhibitors (e.g., allocolchicine, halichondrin M, colchicine, colchicine derivatives, trityl cysteine, vinblastine sulfate, and vincristine sulfate),Plant alkaloids (e.g., actinomycin D, bleomycin, L-asparaginase, idarubicin, vinblastine sulfate, vincristine sulfate, mithramycin, mitomycin, daunorubicin, VP-16-213, VM-26, navelbine, and taxotere), biologics (e.g., alpha interferon, BCG, G-CSF, GM-CSF, and interleukin 2), topoisomerase I inhibitors (e.g., camptothecin, camptothecin derivatives, and morpholinodoxorubicin), topoisomerase II inhibitors (e.g., mitoxantrone, amonafide, m-AMSA, and amphotericin), tetracyclazole derivatives, pyrazoloacridines, bisantrene HCl, daunorubicin, deoxydoxorubicin, menogaril, N,N-dibenzyldaunorubicin, oxanthrazole, rubidazone, VM-26, and VP-16), and synthetic products (e.g., hydroxyurea, procarbazine, o,p'-DDD, dacarbazine, CCNU, BCNU, cis-diamminedichloroplatinum, mitoxantrone, CBDCA, levamisole, hexamethylmelamine, all-trans-retinoic acid, gliadel, and porfimer sodium).

[0059] Therapeutic drugs for treating cancer also include antiproliferative agents, which are compounds that reduce cell proliferation. Antiproliferative agents include alkylating agents, antimetabolites, enzymes, biological response modifiers, various drugs, hormones and antagonists, androgen inhibitors (e.g., flutamide and leuprolide acetate), and antiestrogens (e.g., tamoxifen citrate and its analogs, toremifene, droloxifene, and roloxifene). Additional examples of specific antiproliferative agents include, but are not limited to, levamisole, gallium nitrate, granisetron, sargramostim, strontium chloride, filgrastim, pilocarpine, dexrazoxane, and ondansetron.

[0060] Therapeutic drugs for treating cancer can be administered alone or in combination with other anti-tumor drugs, including cytotoxic / antineoplastic drugs and angiogenesis inhibitors. Cytotoxic / antineoplastic drugs are defined as drugs that attack and kill cancer cells. Some cytotoxic / antineoplastic drugs are alkylating agents that alkylate genetic material in tumor cells, such as cisplatin, cyclophosphamide, nitrogen mustard, trimethylene thiophosphramide, carmustine, busulfan, chlorambucil, belustine, uracil mustard, chlomaphazin, and dacarbazine. Other cytotoxic / antineoplastic drugs are antimetabolites for tumor cells, such as cytosine arabinoside, fluorouracil, methotrexate, mercaptopurine, azathioprim, and procarbazine. Other cytotoxic / antineoplastic agents are antibacterial agents, such as doxorubicin, bleomycin, dactinomycin, daunorubicin, mithramycin, mitomycin, mitomycin C, and daunomycin. Numerous liposomal formulations of these compounds are commercially available. Still other cytotoxic / antineoplastic agents are mitotic inhibitors (vinca alkaloids). These include vincristine, vinblastine, and etoposide. Various cytotoxic / antineoplastic agents include taxol and its derivatives, L-asparaginase, antitumor antibodies, dacarbazine, azacitidine, amsacrine, melphalan, VM-26, ifosfamide, mitoxantrone, and vindesine.

[0061] In some embodiments, the therapeutic drugs described herein include at least one antibacterial agent. The antibacterial agent can be an antifungal agent or an antibacterial agent. In some embodiments, the antibacterial agent is a broad-spectrum antibacterial agent. Suitable broad-spectrum antibacterial agents include, but are not limited to, anti-staphylococcal penicillins such as nafcillin or oxacillin, or first generation cephalosporins such as cefazolin, third generation cephalosporins such as cefotaxime or ceftriaxone, linezoid piperacillin-tazobactam, ampicillin-sulbactam, ticarcillin-clavulanate, doxycycline, minocycline, clindamycin, vancomycin, daptomycin, metronidazole, amoxicillin-clavulanuate, fluoroquinolones such as ciprofloxacin, levofloxacin, moxifloxacin, and trimethoprim-sulfamethoxazole. Antifungal agents include, but are not limited to, lotrimazole, miconazole, ketoconazole, itraconazole and fluconazole, amphotericin B, anidulafungin, and caspofungin.

[0062] In some embodiments, the therapeutic agent is selected from, by way of non-limiting example, at least one nucleotide (e.g., a polynucleotide), at least one carbohydrate, or at least one amino acid (e.g., a peptide). In certain embodiments, the therapeutic agent is a polynucleotide, an oligonucleotide, a gene expression modulator, a knockdown agent, an siRNA, an RNAi agent, a dicer substrate, an miRNA, an shRNA, an antisense oligonucleotide, or an aptamer. In other embodiments, the therapeutic agent is an aiRNA (Asymmetric RNA duplexes mediate RNA interference in mammalian cells. Xiangao Sun, Harry A Rogoff, Chiang J Li Nature Biotechnology 26, 1379-1382 (2008)). In some embodiments, the therapeutic agent is a protein, peptide, dominant-negative protein, enzyme, antibody, or antibody fragment. In some embodiments, the therapeutic agent is a carbohydrate or a small molecule. In some embodiments, the therapeutic agent is a non-biological, synthetic polymer. In some embodiments, the therapeutic drug comprises an autologous or allogeneic stem cell graft, which may or may not be cultured or genetically modified.

[0063] In some embodiments, the therapeutic agent comprises a cell mobilization agent, which acts through one or more mechanisms, including, but not limited to, mobilizing cells from their native environment; preventing inhibition of cell escape; reducing adhesion of cells to their surrounding environment; and modulating neural or cellular controls that determine cell stability, entry, or egress from their environment. In some embodiments, the cell mobilization agent may comprise any suitable composition that can increase cell yield by modulating the neural control of cell mobilization, reducing adhesion to native tissues, either directly or indirectly, or by creating a cell mobilization effect. Cells may be mobilized within the surrounding extracellular environment / matrix, which may include the surrounding vascular space; in the case of bone marrow, this may be within the surrounding capillary beds and sinusoids.

[0064] Non-limiting classes of drugs that can be used include currently known and yet undiscovered classes of known proteins and receptors that control the recruitment of cells out of tissues or the retention of cells within tissues. These can include combinations, such as the inhibition of α9β1 with BOP (an antibody or other small molecule selective inhibitor) and AMD3100 in combination. These can include broad classes, such as: integrin family, such as the VLA-4 molecule inhibitors filategast, UNII-OJY3SK9H5F, BIO5192, and their derivatives; modulation of CXCL12 / CXCR4 interactions, such as the CXCR4 inhibitor plerixafor; modulation of CXCR7 molecules; CXCL12 analogs; modulation of neuronal / stem cell interactions, such as dopamine receptors (1-5 subtypes) and noradrenergic α and β receptors and all such receptors, their precursors and derivatives for catecholamines; modulators or synergists of these receptors. Modulators of neurotransmitter uptake from the nasal / site of action; inhibitors of catecholamine breakdown, such as inhibitors of catechol-O-methyltransferase (COMT) or amination by the monoamine oxidase (MAO) enzyme; modulators of the downstream cascade of catecholamine receptors, such as inhibitors of adenylate cyclase and other phosphoinositide 3-kinase (PI3K) / Akt pathways; agonists and blockers of catecholamine receptors a1, a2, b1, b2, and b3; catecholamine precursors, such as L-phenylalanine, L-tyrosine, and L-DOPA;Dopamine agonists such as aripiprazole, phencyclidine, quinpirole, salvinorin A, apomorphine, bromocriptine (Parlodel), cabergoline (Dostinex), ciladopa, dihydrexidine, dinapsoline, doxanthrine, epicriptine, lisuride, pergolide, piribedil (Pronolan and Trivastal), pramipexole (Mirabex and Siflor), rol), propylnorapomorphine, quinagolide (Norprolac), ropinirole, rotigotine, loxindol, sumanilol, fenoldopam selective for dopamine D1 receptors, cocaine, amphetamine; dopamine reuptake inhibitors such as bupropion, altropane (O-587), amfonelic acid (WIN25978), amineptine (which has a reasonable degree of selectivity for dopamine over norepinephrine reuptake inhibitors), BTCP (GK-13 ), 3C-PEP (remarkable potency and selectivity for dopamine transporters), DBL-583, difluoropine (O-620), GBR-12783, GBR-12953, GBR-13069, GBR-13098, GYKI-52895, Iometopane (β-CIT, RTI-55), methylphenidate, ethylphenidate, modafinil, armodafinil, RTI-229, vanoxerine (GBR-12909), haloperidol, Chlorpromazine, eticlopride, pimozide, chlorpromazine, eticlopride; desipramine and other drugs that inhibit norepinephrine reuptake; DRD1, DRD2, DRD3, DRD4, DRD4 receptor agonists and antagonists such as eticlopride; nicotine; b2-adrenergic agonists such as clenbuterol; α9 integrin agonists; BOP, N-(benzene-sulfonyl)-L-prolyl-LO-(1-pyrrolidinylcarbonyl)tyrosine;VLA-4 antagonists, such as trans-4-[1-[[2-(5-fluoro-2-methylphenylamino)-7-fluoro-6-benzoxazolyl]acetyl]-(5S)-[methoxy(methyl)amino]methyl-(2S)-pyrrolidinylmethoxy]cyclohexanecarboxylic acid, natalizumab, and BIO5192; matrix metalloproteinases and their inducers, such as Me6TREN; prolyl hydroxylase inhibitors, such as dimethyloxalylglycine (DMOG); chemokine GRObeta; sulfated colominic acid; beta-chemokine CCL15; Panax notoginseng saponin (panax notoginseng saponins); VEGF; ALT-1188; P2RY14 agonists, e.g., MRS2690; UDP-glucose; gamma-tocotrienol; TGFβ, TGF-β1, and substance P; modulation of adhesion molecules, e.g., VCAM-1; interactions with integrins, such as VLA-4 (α9β1); G protein-coupled receptors, such as P2Y purinergic receptor-14; S1P-1 modulators, including ATC-128800, SEW2871, GSK2018682, FTY720, MRS2690, and dopamine; various endocrine targets, such as NOTCH proteins (parathyroid hormone); granulocyte colony-stimulating factor (G-CSF) and analogs (filgrastim); pegylated and glycosylated forms of G -CSF; granulocyte-macrophage colony-stimulating factor (GM-CSF); macrophage colony-stimulating factor (M-CSF); tyrosine kinase 3 (FLT-3); ancestim; stem cell factor; AMD3100; TG-0054; KRP203; 4F-benzoyl-TM14003; POL6326; P2G, a mutant protein of SDF-1β; CTCE-0021; CS549; pepducins such as ATI-2341; cytokines (e.g., interleukin-1, interleukin-3, interleukin-6, interleukin-7, interleukin-11, interleukin-12); metalloproteases; serine proteases; cysteine ​​proteases; peptidases; chemokines; etc. Also included are non-selective agents such as heparin, sulfates and derivatives, and amide-type anesthetics such as lidocaine.

[0065] In some embodiments, the therapeutic agent may comprise any suitable composition capable of reducing the sensation of pain, non-limiting examples of which include one or more of lidocaine, prilocaine, tetracaine, benzocaine, procaine, mepivacaine, bupivacaine, etidocaine, tropacocaine, piperocaine, stovaine, cyclomethylcaine, parethoxycaine, dyclonine, falicain, pramoxine, amolanone, phenacaine, diperodone, dibucaine, and the like.

[0066] Vasoactive drugs

[0067] Studies of the effects of vasoactive agents on blood flow through the bone marrow and the vascular resistance of the circulation in the bone marrow have shown that these agents affect the rate at which they reach the systemic circulation. Vasoactive agents, such as those used in combination with anesthetics (e.g., lidocaine) and epinephrine, reduce blood flow to the skin and subcutaneous tissues. Epinephrine reduces blood flow to the skin, thereby slowing the rate at which lidocaine is absorbed into the circulation and allowing larger doses to be administered without systemic toxicity.

[0068] In some embodiments, the present invention provides methods of using vasoactive agents to regulate blood flow. Vasoactive agents may be pharmacologically synergistic or additive for their intended purpose, e.g., alpha and beta adrenergic receptor or selective or non-selective dopamine receptor modulation, and mobilize stem cells. In one embodiment, infusates that cause stem cell release from their niches can be combined with vasoconstrictive substances to reduce blood flow and prevent the loss of mobilized cells to the systemic circulation. Administration of vasoactive agents may thereby maintain cells that have escaped from their niches within the sinusoids and / or arterioles within the bone. In some embodiments, vasodilatory substances may be used to flush out unwanted infusates that have localized within the bone. Administration of vasoactive agents thereby allows for the controlled elimination of procedures in which further maintenance of the infusate in the bone is no longer desired.

[0069] In some embodiments, the vasoactive agent comprises a vasoconstrictor, including, but not limited to, sympathomimetics, specific and non-specific modulators of alpha and beta adrenergic receptors and subtypes, including specific modulation of dopamine subtypes, and modulators of dopamine receptors, direct and indirect agonists and dopamine-releasing agents such as dopamine reuptake inhibitors, methoxamine hydrochloride, epinephrine, norepinephrine, dobutamine, dopamine, milrinone, midodrine hydrochloride, desglimidodrine, and the like.

[0070] In some embodiments, the vasoactive agent comprises a vasodilator, including, but not limited to, alpha or beta adrenergic receptor modulators, muscarine, histamine, dopamine receptor modulators, organic nitrates, isosorbide mononitrate, mononitrates, isosorbide dinitrate, dinitrates, nitroglycerin, trinitrates, minoxidil, sodium nitroprusside, hydralazine hydrochloride, nitric oxide, nicardipine hydrochloride, fenoldopam mesylate, diazoxide, enalaprilat, epoprostenol sodium, prostaglandins, vasoactive intestinal peptide (VIP), substance P, niacin, carbon dioxide, natriuretic peptides, heparin, bradykinin, L-arginine, histamine and histamine agonists, antihistamines, minorulin lactate, bipyridine, dopamine D1-like receptor agonists, stimulants or activators, and the like.

[0071] In some embodiments, vasoconstrictors and vasodilators may be used to regulate intraosseous pressure for the purpose of regulating bone volume, hi some embodiments, vasoconstrictors and vasodilators may be used to regulate intraosseous pressure as a primary endpoint for treating a disease.

[0072] additives

[0073] In various embodiments, the injectable compositions of the present invention may further comprise additional additives, including drugs, pharmaceutical agents, carriers, buffers, adjuvants, dispersants, diluents, etc., depending on the intended use and application. In various embodiments, the additives may be used to adjust certain properties of the composition, including, but not limited to, viscosity, lipophilicity, hydrophobicity, hydrophilicity, pH, polarity, etc.

[0074] Examples of suitable pharmaceutical carriers, excipients and / or diluents are well known in the art and include, but are not limited to, gums, starches (e.g., corn starch, pregelatinized starch), sugars (e.g., lactose, mannitol, sucrose, dextrose), cellulosic materials (e.g., microcrystalline cellulose), acrylates (e.g., polymethylacrylate), calcium carbonate, magnesium oxide, talc, or mixtures thereof.

[0075] Pharmaceutically acceptable carriers for liquid preparations are aqueous or non-aqueous solutions, suspensions, emulsions or oils. Examples of non-aqueous solvents are propylene glycol, polyethylene glycol, and injectable organic esters such as ethyl oleate. Examples of oils are those of animal, vegetable, or synthetic origin, such as peanut oil, soybean oil, olive oil, sunflower oil, turmeric oil, fish liver oil, marine oil, or fats from milk or eggs.

[0076] Aqueous carriers include water, alcoholic / aqueous solutions, emulsions, or suspensions containing saline buffer, e.g., phosphate-buffered saline, water, emulsions, e.g., oil / aqueous emulsions, various types of wetting agents, sterile solutions, etc. Compositions containing such carriers can be formulated using well-known, conventional methods. Suitable carriers can contain any material that, when combined with the biologically active compounds of the present invention, retains biological activity. Preparations for parenteral administration can include sterile aqueous or non-aqueous solutions, suspensions, and emulsions. Examples of non-aqueous solvents are polypropylene glycol, polyethylene glycol, vegetable oils, e.g., olive oil, and injectable organic esters, e.g., ethyl oleate. Aqueous carriers include water, alcoholic / aqueous solutions, emulsions, or suspensions containing saline buffer. Parenteral vehicles can include sodium chloride solution, Ringer's dextrose, dextrose and sodium chloride, lactated Ringer's, or fixed oils. Intravenous vehicles may include fluid and nutrient replenishers, electrolyte replenishers (e.g., those based on Ringer's dextrose), etc. Preservatives and other additives may also be present including, for example, antimicrobial agents, antioxidants, chelating agents, and inert gases and the like; in addition, the pharmaceutical compositions of the invention may include a proteinaceous carrier, such as serum albumin or immunoglobulin of human origin.

[0077] The compositions provided herein can also be administered as controlled-release compositions, i.e., compositions in which the active ingredient is released over a period of time after administration. Controlled-release or sustained-release compositions include formulation in lipophilic depots (e.g., fatty acids, waxes, oils). In another embodiment, the compositions are immediate-release compositions, i.e., compositions in which all of the active ingredient is released immediately after administration.

[0078] As used herein, "additional ingredients" includes, but is not limited to, one or more of the following: excipients, surfactants, dispersing agents, inert diluents, granulating and disintegrating agents; binders; lubricating agents; sweetening agents; flavoring agents; coloring agents; preservatives; physiologically degradable compositions such as gelatin; aqueous vehicles and solvents; oily vehicles and solvents; suspending agents; dispersing or wetting agents; emulsifying agents; analgesics; buffers; salts; thickening agents; fillers; emulsifiers; antioxidants; antibiotics; antifungal agents; stabilizers; and pharmaceutically acceptable polymeric or hydrophobic materials.

[0079] Administration method

[0080] As described elsewhere herein, the methods of the present invention adjust the method of administration to control the release rate and intraosseous distribution of the injectate. In some embodiments, the methods adjust the method of injectate administration for maximum retention in the bone.

[0081] 2, an exemplary method 200 is shown for adjusting the release rate and intra-bone distribution of an injectate. Method 200 begins with step 202, in which an amount of at least one injectate containing at least one therapeutic drug is selected. In step 204, a concentration of the at least one therapeutic drug is optionally selected. In step 206, an infusion rate of the at least one injectate is optionally selected. In step 208, at least one bone site for injecting the at least one injectate is optionally selected. In step 210, the amount of the at least one injectate is injected into the at least one bone site. In step 212, the at least one injectate is maintained in a localized state within the at least one bone site.

[0082] In some embodiments, the methods can be described as maintaining a percentage of the injectate volume at the injection site or within the target bone. For example, the methods can maintain at least 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, or 95% of the injectate volume. In some embodiments, the maintenance percentage can be described as a function of time. For example, the methods can maintain at least 15% of the injectate volume for at least about 0.5 minutes, 1 minute, 2 minutes, 5 minutes, 10 minutes, 30 minutes, 60 minutes, 120 minutes, 24 hours, or 48 hours after injection.

[0083] In some embodiments, the methods of the present invention include a step for selecting the amount and concentration of the injectate, optionally including a therapeutic drug, a vasoactive agent, and combinations thereof. The amount and concentration can be selected based on certain factors, such as the effective amount of the vasoactive agent, the effective amount of the therapeutic drug, and the type and size of the bone associated with the bone site. In some embodiments, the amount and concentration of the vasoactive agent can be selected to reduce blood flow at the bone site by about 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, or 95%. The therapeutically effective amount and concentration of the therapeutic drug will depend on the type of therapeutic drug, the condition being treated or prevented, etc. In some embodiments, the therapeutic drug is present at greater than about 0%, greater than about 5%, greater than about 10%, greater than about 15%, greater than about 20%, greater than about 25%, greater than about 30%, greater than about 35%, greater than about 40%, greater than about 45%, greater than about 50%, greater than about 55%, greater than about 60%, greater than about 65%, greater than about 70%, greater than about 75%, greater than about 80%, greater than about 85%, greater than about 90%, greater than about 95% by weight. Typical volumes that can be injected into bone can range between about 0.1 μL, 10 μL, 50 μL, 100 μL, 500 μL, 1 mL, 2 mL, 3 mL, 4 mL, 5 mL, 10 mL, 20 mL, 50 mL, etc. In situations where the bone is irrigated multiple times with successive treatments, the amount may be described as a cumulative amount, for example, between about 100 mL and 1000 mL. It should be understood that a treatment or procedure may require a larger amount of injectate, in which case the total amount may be injected in smaller increments.

[0084] In some embodiments, the methods of the present invention optionally include a step for selecting an infusion rate for the infusate. The infusion rate can be about 0.1 μL / s, 1 μL / s, 10 μL / s, 50 μL / s, 0.1 mL / s, 0.2 mL / s, 0.3 mL / s, 0.4 mL / s, 0.5 mL / s, 0.6 mL / s, 0.7 mL / s, 0.8 mL / s, 0.9 mL / s, 1 mL / s, 2 mL / s, 3 mL / s, 4 mL / s, 5 mL / s, 6 mL / s, 7 mL / s, 8 mL / s, 9 mL / s, 10 mL / s, etc. In some embodiments, the vasoactive agent and the therapeutic agent are infused at different rates. In some embodiments, the treatment rate is constant. In some embodiments, the treatment rate is variable, such that the infusion rate increases or decreases over time. For example, the infusion rate can be varied over a range of between about 0.1 μL / s to 0.5 μL / s, 0.5 μL / s to 1 μL / s, 1 μL / s to 5 μL / s, 1 μL / s to 0.1 mL / s, 0.1 mL / s to 0.5 mL / s, 0.5 mL / s to 1 mL / s, 1 mL / s to 5 mL / s, 1 mL / s to 10 mL / s, etc. It should be understood that the infusion rate will vary depending on the viscosity of the injectate, the volume of the injectate, the density of the bone, etc. In some embodiments, the viscosity of the injectate can be varied for both therapeutic and area dose effects. Viscosity can range from about 5 to 800 cP (800 cP is approximately twice the reported viscosity of bone marrow in some mammals).

[0085] In some embodiments, the methods of the present invention optionally include a procedure for selecting the anatomical location of the injectate. For example, an injectate injected near the bone's outflow veins is more likely to be released into the systemic circulation at a faster rate than an injectate injected away from the outflow veins. A vasoactive agent injected near the bone vasculature is more likely to regulate intra- and extra-bone circulation than an injectate injected away from the bone vasculature. In some embodiments, the injectate can be injected at a bone site away from the outflow veins. In some embodiments, the vasoactive agent can be injected at a site closer to the outflow veins than the therapeutic drug to limit the effect of the vasoactive agent on blood outflow / inflow. In some embodiments, the vasoactive agent can be injected into tissue surrounding the bone site targeted for therapeutic drug infusion. In some embodiments, the vasoactive agent can be administered systemically prior to injection of the therapeutic drug into the bone site. In some embodiments, the vasoactive agent can be placed in the artery-feeding bone.

[0086] In some embodiments, the method techniques are enhanced by using pressure-based techniques. For example, as a volume of injectate is injected into a first bone portion, one or more volumes of aspirate are extracted from one or more additional bone portions. A pressure gradient is thereby created across the vasculature or bone region between the first bone portion of injection and one or more additional bone portions of extraction, where the first bone portion of injection has a higher pressure and the one or more additional bone portions each have a lower pressure than the first bone portion, causing the aspirate to move through the one or more additional bone portions. The one or more volumes of aspirate may contain any suitable or desired material, such as injectate, bone, blood, cells, bone marrow, etc. The timing of injection of injectate and removal of material may be simultaneous or staggered. In some embodiments, the injection and removal may be performed using a single device having two lumens, a first lumen for injection of injectate and a second lumen for material removal. Material removal may thereby be performed at the same site as the injection site. In some embodiments, the injection and removal may be performed using two or more separate devices, whereby the injectate is injected at a first site and material is removed from one or more additional sites. The one or more additional sites can be immediately adjacent to each other or separated from the first site by a small distance, e.g., between about 5 mm and about 50 mm. In some embodiments, the one or more additional sites are separated from the first site by about 50 mm to about 500 mm. In exemplary embodiments, simultaneous administration of injectate at one site of the bone and aspiration of material at another site of the bone expands the area of ​​injectate beyond that reachable with a single needle by creating a lower pressure differential than the venous system, which creates preferential flow to the aspiration needle compared to the venous outlet, and by creating a preferential negative pressure that expels injectate across the bone region into the aspiration needle rather than being forced into the venous system by a single pressure head. In some embodiments, the viscosity of the injectate can be selected for optimal in situ replacement of bone marrow within the bone sinusoids, similar to the manner in which injectates used for extracting oil from oil sands are tailored for optimal oil replacement. Bone contents, including bone marrow, cells, infectious particles, and tumors, are transferred to a needle or other device that creates negative pressure for collection.

[0087] In some embodiments, the steps of the method are augmented using mechanical techniques. For example, the method may employ a first device that distributes infusate to multiple locations, e.g., multiple sequential depths, either simultaneously or sequentially. The first device may be used in combination with a second device that aspirates by matching negative pressure at multiple different locations complementary to the multiple locations of the first device. In some embodiments, the first device, the second device, or both may include multiple openings through which infusate or aspirate may pass. The multiple openings may be individually isolated to selectively distribute infusate at various infusion sites and depths and withdraw aspirate at various desired aspiration sites and depths, thereby maintaining physiological pressure and "pulling" the liquid across the aspiration area. The procedure may be repeated at various locations and depths until the entire aspiration area between the first and second devices is administered or collected.

[0088] In one embodiment, methods and agents may be combined to optimize their effectiveness. The bone target site can be flanked by at least one dual-lumen needle, with one lumen exiting the distal side of the needle and the other lumen adjacent to one or more side holes. For example, as depicted in FIG. 3 herein (and illustrated in FIGS. 4-6), an exemplary method 300 is shown. Method 300 is advantageous for separating the agents administered through each lumen of the needle and localizing each agent. For example, if vasoconstriction is not needed throughout the entire bone, but only within the draining vein, separate lumen openings can be used to administer a vasoconstrictor agent to reduce blood, cells, or release of the therapeutic agent or agents from the bone, while leaving the treatment area without vasoconstriction, which may impede the flow of therapeutic agents within the treatment area of ​​the bone.

[0089] Method 300 begins with step 302, in which at least one double-lumen needle is provided, the double-lumen needle having a first lumen terminating in a distal opening and a second lumen with at least one side port proximate the opening of the first lumen (shown in FIG. 4). In step 304, the at least one double-lumen needle is inserted into a target bone site having a delivery area and a vasoconstriction area, with the distal opening of the first lumen positioned toward the vasoconstriction area and the at least one side port of the second lumen positioned toward the delivery area (shown in FIG. 5, in which the target bone site is the iliac crest). In step 306, a vasoconstrictor is delivered through the first lumen and the opening, such that the vasoconstrictor localizes in the vasoconstriction area and reduces blood, cell, or infusate shedding from the delivery area. In step 308, one or more injectates are administered through the second lumen and the at least one side opening such that the one or more injectates are localized in the administration area (shown in FIG. 6).

[0090] In some embodiments, the same double-lumen needle is used to aspirate, extract, and collect tissue (e.g., blood and cells), e.g., through the at least one side port of the second lumen. In some embodiments, one or more additional needles, each having at least one lumen, may be provided. For example, the additional needles may be single-lumen or double-lumen needles having first and second lumens as described above. In some embodiments, the one or more additional double-lumen needles each administer a vasoconstrictor through a distal opening of the first lumen to increase coverage of the vasoconstrictor. The one or more additional needles may include, but are not limited to, administering additional infusate, extracting and collecting tissue (e.g., blood and cells) across a region via a pressure gradient, passively or actively releasing fluid pressure within a target bone site for pain relief, and combinations thereof, via the corresponding second lumen of the single or respective double-lumen needle. An exemplary application of the described method includes administering drugs that result in the release of cells out of their niche within the sinusoids and using separate drugs through separate lumens to retain those cells within the bone by vasoconstriction.

[0091] In some embodiments, the steps of the method are enhanced using physical or electrical techniques. For example, physical methods of enhancing injectate release and intraosseous distribution can be achieved by applying vibrations to the bone, e.g., by ultrasound or massage. Physical methods can also include restricting blood flow, e.g., by applying a temporary cuff, tourniquet, or cold pack to tissue in the vicinity of the bone. As described elsewhere herein, physical methods can also include forced injectate outflow procedures to prepare the injection site. In some embodiments, the injectate can include charged or magnetic particles so that injectate distribution can be controlled through the application of an electric or magnetic field. In some embodiments, the injectate modulates the pH of the intrathecal cavity. In some embodiments, the injectate includes an anticoagulant.

[0092] In some embodiments, various approaches to enhancing the procedure of the present methods are used in combination, including, but not limited to, pharmacological approaches, pressure-based approaches, mechanical approaches, and electrical approaches.

[0093] The various methods and procedures for adjusting the mode of infusion administration are similarly applicable to fluid aspiration, if the treatment or procedure requires it. Each pharmacologically active therapeutic drug has a time curve for its peak effect. In some embodiments, fluid aspiration can be performed after the peak effect has occurred. For example, for the purpose of enhancing stem cell recovery, during bone marrow aspiration, stem cells can be collected at 0, 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, or more than 90 minutes. [Example]

[0094] The present invention is further described in detail in the following examples by reference. These examples are provided for illustrative purposes only and are not intended to be limiting unless specifically stated. Therefore, the present invention should not be construed as being limited to the following examples in any way, but rather as encompassing any and all variations that may be evident as a result of the teachings provided herein.

[0095] Example 1: Maximizing distribution and minimizing spillage of fluid injected into bone

[0096] The results described herein demonstrate that vasoactive agents promote localized retention of injected liquid medication within bone. The injection rate and injection site also promote localized retention of injected liquid medication within bone. Anatomical location influences the distribution and duration of injectate within bone. Also, the viscosity of the injected iodinated contrast agent shown does not fill the entire bone, and therefore a second needle may be placed to administer liquid or provide negative pressure for increased distribution.

[0097] 7 is a fluoroscopic image of contrast agent administered into a subject's bone at a maximum distribution rate of 0.5 mL / sec. The administration rate combined with the viscosity shown does not adequately displace the current blood volume within the bone sinusoids.

[0098] Figure 8 shows a fluoroscopic image of contrast agent being administered into a subject's bone at a maximum distribution rate of 10 mL / sec. The administration rate is so fast that the contrast agent immediately enters the bloodstream without being retained within the bone. This can be improved or eliminated by additional negative pressure from the placement of a second device in the bone distant from the first site.

[0099] Figure 9 shows a fluoroscopic image of contrast agent administered into the bone of a subject at a maximum distribution rate of 5 mL / sec. Noteworthy is the relatively small bone perfusion area due to the constricted intraosseous draining veins and proximity to the central draining vein.

[0100] Figure 10 shows a fluoroscopic image of contrast agent administered without epinephrine into the bone of a subject at a maximum distribution rate of 5 mL / sec. The injection distance from the central draining vein is maximized. Noteworthy is the larger area of ​​drug opacification.

[0101] Figure 11 shows a fluoroscopic image of contrast agent co-administered with epinephrine into the bone of a subject at a maximum distribution rate of 5 mL / sec. The clearance rate of the contrast agent was decreased, and the area of ​​opacification increased compared to placement of the device adjacent to the draining vein. However, the area of ​​opacification was not as extensive compared to injection prior to epinephrine injection, which may be due to a constricted vein restricting lateral flow within the bone. This may be eliminated by a second device in a different location or by administration of a vasoconstrictor to a site adjacent to the bone's draining vein.

[0102] Figure 12 shows a fluoroscopic image of contrast agent administered with epinephrine into the subject's left bone at a maximum distribution rate of 5 mL / sec 20 minutes after injection. The contrast agent with epinephrine remains in a smaller perfusion area.

[0103] The disclosures of each and every patent, patent application, and publication cited herein are incorporated herein by reference in their entirety. Because the invention has been disclosed by reference to certain embodiments, it will be apparent that other embodiments and variations of the invention may be devised by those skilled in the art without departing from the true spirit and scope of the invention as claimed. The appended claims are intended to be construed to include all such embodiments and equivalent variations.

Claims

1. 1. A method for modulating the release rate and intraosseous distribution of an injectate, comprising: providing a quantity of at least one injectate containing at least one therapeutic agent; injecting a quantity of the at least one injectant into the first bone portion via the first lumen; and maintaining said at least one injectate volume within said first bone portion in a localized state such that at least 15% to 95% of said volume is retained for at least 5 minutes to 2 hours; A method comprising the steps of:

2. 10. The method of claim 1, wherein the at least one injectate further comprises at least one vasoactive agent.

3. 2. The method of claim 1, wherein the injecting step is performed after the step of administering at least one first injectate at a pressure or speed such that connections between the peripheral veins of the first bone portion are opened through incomplete or fragile sinusoidal walls.

4. 4. The method of claim 3, wherein the pressure is selected from the group consisting of about 10 psi, 20 psi, 30 psi, 40 psi, 50 psi, 60 psi, 70 psi, 80 psi, 90 psi, 100 psi, 200 psi, 300 psi, 400 psi, 500 psi, 600 psi, 700 psi, 800 psi, 900 psi, and 1000 psi.

5. 4. The method of claim 3, wherein the rate is selected from the group consisting of about 0.1 μL / s, 1 μL / s, 10 μL / s, 50 μL / s, 0.1 mL / s, 0.2 mL / s, 0.3 mL / s, 0.4 mL / s, 0.5 mL / s, 0.6 mL / s, 0.7 mL / s, 0.8 mL / s, 0.9 mL / s, 1 mL / s, 2 mL / s, 3 mL / s, 4 mL / s, 5 mL / s, 6 mL / s, 7 mL / s, 8 mL / s, 9 mL / s, and 10 mL / s.

6. 10. The method of claim 1, wherein an amount of at least one vasoactive agent is infused prior to injecting the amount of the at least one infusate.

7. 10. The method of claim 1, wherein an amount of at least one vasoactive agent is infused after injecting the amount of the at least one infusate.

8. 10. The method of claim 1, wherein the amount of the at least one vasoactive agent is infused simultaneously with the amount of the at least one infusate.

9. 10. The method of claim 1, wherein the amount of at least one vasoactive agent is injected proximate to a bone draining vein.

10. The method of claim 1 , wherein the first bone portion is located non-adjacent to a bone draining vein.

11. 10. The method of any one of claims 2 or 6-9, wherein the at least one vasoactive agent is a vasoconstrictor selected from the group consisting of a sympathomimetic agent, methoxamine hydrochloride, epinephrine, dobutamine, dopamine, norepinephrine, milrinone, midodrine hydrochloride, desglimidodrine, and an alpha receptor agonist, stimulator, or activator.

12. 10. The method of any one of claims 2 or 6-9, wherein the at least one vasoactive agent is a vasodilator selected from the group consisting of alpha or beta adrenergic receptor modulators, muscarine, histamine, dopamine receptor modulators, organic nitrates, isosorbide mononitrate, mononitrates, isosorbide dinitrate, dinitrates, nitroglycerin, trinitrates, minoxidil, sodium nitroprusside, hydralazine hydrochloride, nitric oxide, nicardipine hydrochloride, fenoldopam mesylate, diazoxide, enalaprilat, epoprostenol sodium, prostaglandins, milrinone lactate, bipyridine, and dopamine D1-like receptor agonists, stimulators, or activators.

13. 10. The method of claim 1, wherein the at least one therapeutic agent is selected from the group consisting of antineoplastic agents, antiproliferative agents, antiangiogenic agents, cytotoxic agents, antimicrobial agents (including antibacterial and antifungal agents), cell mobilizing agents, and analgesic agents.

14. 10. The method of claim 1, comprising the step of selecting an infusion rate of the at least one infusate from the group consisting of about 0.1 μL / s, 1 μL / s, 10 μL / s, 50 μL / s, 0.1 mL / s, 0.2 mL / s, 0.3 mL / s, 0.4 mL / s, 0.5 mL / s, 0.6 mL / s, 0.7 mL / s, 0.8 mL / s, 0.9 mL / s, 1 mL / s, 2 mL / s, 3 mL / s, 4 mL / s, 5 mL / s, 6 mL / s, 7 mL / s, 8 mL / s, 9 mL / s, and 10 mL / s.

15. 10. The method of claim 1, wherein the volume of the at least one injectate is selected from the group consisting of about 0.1 μL, 1 μL, 10 μL, 50 μL, 100 μL, 500 μL, 1 mL, 2 mL, 3 mL, 4 mL, 5 mL, and 10 mL.

16. 10. The method of any one of claims 2 or 6-9, wherein the amount of the at least one vasoactive agent is selected to reduce blood flow at the bone by about 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, or 95%.

17. 10. The method of claim 1, wherein the amount of the at least one therapeutic agent has a concentration selected from the group consisting of greater than about 0% by weight, greater than about 5% by weight, greater than about 10% by weight, greater than about 15% by weight, greater than about 20% by weight, greater than about 25% by weight, greater than about 30% by weight, greater than about 35% by weight, greater than about 40% by weight, greater than about 45% by weight, greater than about 50% by weight, greater than about 55% by weight, greater than about 60% by weight, greater than about 65% by weight, greater than about 70% by weight, greater than about 75% by weight, greater than about 80% by weight, greater than about 85% by weight, greater than about 90% by weight, or greater than about 95% by weight.

18. 2. The method of claim 1, further comprising, after the step of maintaining the at least one injectate volume within the first bone portion in a localized state, extracting an aspirate volume from one or more additional bone portions via one or more additional lumens, such that a pressure gradient is formed across a region of vasculature or bone between the first bone portion having a first pressure and the one or more additional bone portions each having a pressure lower than the first pressure.

19. 20. The method of claim 18, wherein the first lumen and the one or more additional lumens are disposed within a single device.

20. 20. The method of claim 18, wherein the first lumen is disposed within a first device and the one or more additional lumens are disposed within one or more additional devices.

21. 20. The method of claim 18, wherein the one or more additional bone portions are each spaced between about 5 mm and about 500 mm from the first bone portion.

22. 20. The method of claim 18, wherein the injecting and extracting steps are performed at sequential depths in their corresponding bone portions.

23. 1. A method of increasing aspirate yield from a bone portion, comprising: providing a quantity of at least one injectate; injecting a quantity of the at least one injectant into the first bone portion through the first lumen; and extracting a volume of aspirate from one or more additional bone portions via one or more additional lumens; wherein the first bone portion and the second bone portion are located between about 5 and 500 mm from each other; A method in which a pressure gradient is created across a region of vasculature or bone between the first bone portion having a first pressure and one or more additional bone portions each having a pressure lower than the first pressure.

24. 24. The method of claim 23, wherein the steps of injecting and extracting are performed simultaneously.

25. 24. The method of claim 23, wherein the extracting step is performed consecutively after the injecting step.

26. 26. The method of any one of claims 23 to 25, wherein the injecting and extracting steps are performed at sequential depths in their corresponding bone portions.

27. 26. The method of any one of claims 23 to 25, wherein the first lumen and the one or more additional lumens are disposed within a single device.

28. 26. The method of any one of claims 23 to 25, wherein the first lumen is disposed in a first device and the one or more additional lumens are disposed in one or more additional devices.

29. 26. The method of any one of claims 23-25, further comprising the step of selecting the positive infusion pressure and the negative extraction pressure from the group consisting of about 10 psi, 20 psi, 30 psi, 40 psi, 50 psi, 60 psi, 70 psi, 80 psi, 90 psi, 100 psi, 200 psi, 300 psi, 400 psi, 500 psi, 600 psi, 700 psi, 800 psi, 900 psi, and 1000 psi.

30. 26. The method of any one of claims 23-25, further comprising the step of selecting the infusion and extraction rates from the group consisting of about 0.1 μL / s, 1 μL / s, 10 μL / s, 50 μL / s, 0.1 mL / s, 0.2 mL / s, 0.3 mL / s, 0.4 mL / s, 0.5 mL / s, 0.6 mL / s, 0.7 mL / s, 0.8 mL / s, 0.9 mL / s, 1 mL / s, 2 mL / s, 3 mL / s, 4 mL / s, 5 mL / s, 6 mL / s, 7 mL / s, 8 mL / s, 9 mL / s, and 10 mL / s.

31. 26. The method of any one of claims 23-25, wherein the volume of the at least one injectate and the volume of the aspirate are each selected from the group consisting of about 0.1 μL, 1 μL, 10 μL, 50 μL, 100 μL, 500 μL, 1 mL, 2 mL, 3 mL, 4 mL, 5 mL, and 10 mL.

32. 26. The method of any one of claims 23 to 25, wherein the amount of aspirant is a percentage of the amount of the at least one injectant, said percentage being selected from the group consisting of 50%, 60%, 70%, 80%, 90%, and 100%.

33. 26. The method of any one of claims 23 to 25, wherein the at least one injectate quantity comprises a liquid.

34. 26. The method of any one of claims 23 to 25, wherein the at least one injectate volume comprises at least one fluid agent selected from vasoactive agents, anti-tumor agents, anti-proliferative agents, anti-angiogenic agents, cytotoxic agents, anti-microbial agents (including anti-bacterial and anti-fungal agents), cell mobilizing agents, and analgesic agents.

35. 35. The method of claim 34, wherein the vasoactive agent is a vasoconstrictor selected from the group consisting of sympathomimetics, methoxamine hydrochloride, epinephrine, dobutamine, dopamine, norepinephrine, milrinone, midodrine hydrochloride, desglimidodrine, and alpha receptor agonists, stimulators, or activators.

36. 35. The method of claim 34, wherein the vasoactive agent is a vasodilator selected from the group consisting of alpha or beta adrenergic receptor modulators, muscarine, histamine, dopamine receptor modulators, organic nitrates, isosorbide mononitrate, mononitrates, isosorbide dinitrate, dinitrates, nitroglycerin, trinitrates, minoxidil, sodium nitroprusside, hydralazine hydrochloride, nitric oxide, nicardipine hydrochloride, fenoldopam mesylate, diazoxide, enalaprilat, epoprostenol sodium, prostaglandins, milrinone lactate, bipyridine, and dopamine D1-like receptor agonists, stimulants, or activators.

37. 35. The method of claim 34, wherein the at least one fluid agent has a concentration selected from the group consisting of greater than about 0% by weight, greater than about 5% by weight, greater than about 10% by weight, greater than about 15% by weight, greater than about 20% by weight, greater than about 25% by weight, greater than about 30% by weight, greater than about 35% by weight, greater than about 40% by weight, greater than about 45% by weight, greater than about 50% by weight, greater than about 55% by weight, greater than about 60% by weight, greater than about 65% by weight, greater than about 70% by weight, greater than about 75% by weight, greater than about 80% by weight, greater than about 85% by weight, greater than about 90% by weight, or greater than about 95% by weight.

38. 1. A method of localizing an injectate within a bone, comprising: providing at least one dual lumen needle having a first lumen terminating in a distal opening and a second lumen having at least one side port adjacent the opening of the first lumen; inserting the at least one double lumen needle into a target bone site having a delivery area and a vasoconstriction area such that the distal opening of the first lumen is positioned toward the vasoconstriction area and the at least one side port of the second lumen is positioned toward the delivery area; administering a vasoconstrictor through the first lumen and the opening such that the vasoconstrictor localizes within the vasoconstricted area and reduces blood, cell, or infusate shedding from the area of ​​administration; and administering one or more injectates through the second lumen and through the at least one side opening such that the one or more injectates are localized at the administration area. A method comprising the steps of:

39. 39. The method of claim 38, further comprising the step of extracting tissue with the at least one double lumen needle through the at least one side port of the second lumen.

40. 39. The method of claim 38, wherein at least one additional needle is provided, said needle having at least one lumen.

41. 41. The method of claim 40, wherein the at least one additional needle is a single lumen needle.

42. 42. The method of claim 41, further comprising the steps of inserting the at least one additional needle into at least one bone portion adjacent the target bone portion and aspirating using the at least one additional needle.

43. 41. The method of claim 40, wherein the at least one additional needle is a double lumen needle having a first lumen terminating in a distal opening and a second lumen having at least one side port adjacent the opening of the first lumen.

44. 44. The method of claim 43, further comprising the steps of inserting the at least one additional double-lumen needle into at least one bone portion adjacent to the target bone portion, and administering a vasoconstrictor using the first lumen and a distal opening of the at least one additional double-lumen needle.

45. 44. The method of claim 43, further comprising the steps of inserting the at least one additional double lumen needle into at least one bone portion adjacent to the target bone portion, and aspirating using the lumen of the at least one additional double lumen needle.

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