Aspiration device

Abstract

To provide improved devices for biopsy, aspiration and stem cell acquisition, and methods of using the same, the devices balancing aspiration with concurrent infusion to manage changes in pressure at the site of biopsy, where the present invention can be adapted for any biopsy, aspiration or cell harvest procedure, including adipose tissue aspiration and bone marrow aspiration (BMA).SOLUTION: In particular, the present invention limits patient pain, prevents blood contamination, and increases cell mobilization, such as improved stem cell yields with intraosseous (IO) pharmacological mobilization of stem cells during a BMA procedure, and with improved stem cell yields using pharmacological mobilization of stem cells from fat. The pharmacological mobilization of cells allows the harvest of cells from a biopsy that is many times larger than existing methods.SELECTED DRAWING: Figure 1

Description

Technical Field

[0001] Cross - reference to related applications This application claims priority to U.S. Provisional Application No. 62 / 411,780, filed October 24, 2016, and U.S. Provisional Application No. 62 / 504,090, filed May 10, 2017, the contents of which are hereby incorporated by reference in their entirety.

Background Art

[0002] Stem cells are harvested through a number of routes. Among these, the most common are from adipose tissue and bone marrow. Bone marrow aspiration (BMA) is performed for bone marrow transplantation, diagnosis of blood cancers, and regenerative medicine purposes. In the United States, approximately 700,000 BMA procedures are performed annually for blood cancer diagnosis, and an estimated 500,000 - 1,000,000 BMA procedures are performed each year for stem cell therapy and orthopedic procedures. In the field of cancer, BMA is used for the diagnosis, monitoring, and research of hematological malignancies, but sampling limitations restrict up to 27% of samples in diagnosis. A sufficient number of cancer cells from BMA are required to establish in vitro colonies and patient - specific in vivo chemotherapy testing in the "avatar" model of humanized mice. However, for xenografting, at least 3×10 7 cells from BMA are required, and in some diseases, only half of the BMA reaches this threshold (Rongvaux A et al., Nature biotechnology 32.4 (2014): 364 - 372).

[0003] Various types of stem cells can be collected from bone marrow-derived hematopoietic stem cells (HSCs), epithelial stem cells (ESCs), mesenchymal stem cells (MSCs), and others. As of the time of writing, the field of regenerative medicine is rapidly advancing with hundreds of clinical trials registered with the FDA. This field has numerous other novel orthopedic applications (Lodi D et al., Journal of Experimental & Clinical Cancer Research 30.1 (2011): 9; Cavallo C et al., Journal of biological regulators and homeostatic agents 30.2 (2016): 409; Chahla J et al., Orthopaedic journal of sports medicine 4.1 (2016): 1-8; Cruz-Pardos A et al., Hip international: the journal of clinical and experimental research on hip pathology and therapy 26 (2016): 432-7; Gianakos A et al., Journal of orthopaedic trauma 30.1 (2016): 1-9; Hernigou P et al., International Orthopaedics 41.1 (2017): 127-132; Holton J et al., Orthopedic reviews 8.3 (2016): 6659; Khafagy WW et al., Colorectal Disease 19.1 (2017):O66-O74; Kim SJ et al., Cell Transplantation (2017); Lanham NS et al., Foot & Ankle Specialist (2016): 1938640016679697; Prologo JD et al., Clinical Radiology 71 (2016): 307-11) discuss 3D-printed organs and joint surfaces, bone regeneration (Zigdon-Giladi H et al., World Journal of Stem Cells 7.3 (2015):630), and intervertebral disc regeneration (Vadala G et al., World Journal of Stem Cells 8.5 (2016): 185; Vadala G et al., Journal of biological regulators and homeostatic agents 30.4 Suppl 1 (2016): 173) are expected to provide patients with benefits. Stem cells can "enter" the site of injury / degeneration and, due to their anti-inflammatory properties, can treat autoimmune diseases (Ullah I et al., Bioscience reports 35.2(2015):e00191). There are several ongoing clinical trials regarding the use of stem cells for autoimmune diseases (Ullah I et al., Bioscience reports 35.2(2015):e00191). For example, one application involves spinal fusion, where stem cells aspirated from bone marrow are added to a synthetic bone graft to rapidly form a new fused vertebral body together (Clough BH et al., The Spine Journal 17.3(2017):418-430).

[0004] Common problems in current BMA procedures include a low number of desired cell types, hemodilution, and pain. Bone is a large venous space, and venous blood continuously immerses parenchyma-attached stem cells in the parenchyma tissue. The vacuum created by syringe aspiration creates a pressure gradient from the syringe to the bone and then to the peripheral blood. Because the blood has low viscosity, it preferentially flows into the bone marrow (Gurkan UA et al., Annals of biedicalical engineering 36.12(2008):1978-1991). Numerous studies have demonstrated that only 1–2 mL of bone marrow can be reliably obtained from a single site, and that the larger the volume aspirated from a single site, the lower the yield (Batinic D et al., Bone marrow transplantation 6.2 (1990): 103-107; Muschler GF et al., J Bone Joint Surg Am 79.11 (1997): 1699-1709; Bacigalupo A et al., Bone marrow transplantation 9.6 (1992): 467-470; Helgestad J et al., Pediatric blood & cancer 57.2 (2011): 224-226; Li J et al., Chinese Journal of Cancer Research 23.1 (2011): 43-48; Wang TF et al., Biology of Blood and Marrow Transplantation 17.3 (2011): 351-355; Loken MR et al., Cytometry Part B: Clinical Cytometry 76.1 (2009): 27-36; Fennema EM et al., Acta orthopaedica 80.5 (2009): 618-621; Riley RS et al., Journal of clinical laboratory analysis 18.2 (2004): 70-90).Other experiments using radiolabeled red blood cells suggest that stem cells are aspirated within the blood fluid volume (Holdrinet RSG et al., Experimental hematology 8(1980):103-7). In summary, current bone marrow aspiration yields a small number of mesenchymal stem cells, 1 × 10⁶. 5 Only about 1 to 10 cells, or 0.0001% to 0.01% of all nucleated cells in the bone marrow, can be obtained per individual cell.

[0005] Hemodilution in cancer studies, even with small amounts of typical cancer assessment (7 ml), can result in aspicular hemodiluted samples that limit diagnosis. A recent study in tertiary care facilities quantified the proportion of uncorrected aspirations by advanced pathological and histological techniques in a retrospective review of bone marrow aspiration and biopsy (BMAB). 350 patients underwent at least one inadequate aspiration and at least one subsequent aspiration for comparison. Of the 1250 aspirations in these 350 patients, 470 (27%) were limited, and 58% of these appeared clinically significant in that they were not corrected by core flow cytometry or cytogenetics. 7.7% required repeat biopsy, and 4% had important diagnoses missed. An additional retrospective review of donation registries for patients with myelodysplastic syndromes (MDS) was conducted to compare cell counts by disease subtype classification. Xenotransplantation involved 3 × 10⁶ 7 A rough cutoff of individual cells is required. Only 52% of these procedures yielded enough cells for transplantation.

[0006] In orthopedic surgery, hemodilution affects the success of procedures because the procedure largely depends on the number of stem cells obtained during bone marrow aspiration (BMA). Each patient has a different stem cell profile, and elderly and frail patients generally have fewer stem cells available in the bone marrow and other tissues. Often, up to 60 mL is collected from a single site, resulting in a large volume of peripheral blood. Due to the difficulties arising from collecting cells from bone marrow, there is interest in obtaining stem cells from fat. Adipose tissue is an attractive source of MSCs for stem cell therapy because it is readily available in sufficient quantities through minimally invasive procedures. Furthermore, adipose tissue contains more MSCs than bone marrow (approximately 100,000 MSCs per gram of fat). However, this sampling method is still limited by the amount of fat that can be directly aspirated with the device.

[0007] BMA is the cause of moderate to severe pain in up to 87% of patients (Vanhelleputte P et al., Journal of pain and symptom management 26.3 (2003): 860-866; Mainwaring CJ et al., International Journal of Laboratory Hematology 18.4 (1996): 285-288; Vigneault L et al., Canadian Journal of Anesthesia 58.1 (2011): 22-37). Pain is an obstacle in hematological cancer trials and a significant cause of decreased bone marrow registration, thus limiting bone marrow donation by healthy individuals (Switzer GE et al., Bone marrow transplantation 24.3 (1999); Johansen KA et al., Transfusion Medicine 18.4 (2008): 250-259; Hyde MK et al., Psychology, health & medicine 19.1 (2014): 115-125). It also increases the cost to hospitals in providing sedation procedures to treat pain. [Overview of the Initiative] [Problems that the invention aims to solve]

[0008] Therefore, there is a need in this field to increase the yield of stem cells from both fat and bone marrow, and to reduce the pain of bone marrow aspiration. This invention addresses this need. [Means for solving the problem]

[0009] In one aspect, the present invention relates to a suction device comprising at least one first elongated member having a lumen extending between a proximal opening and a distal tip opening, and a second elongated member having a lumen extending between a proximal opening and a distal tip opening, and at least one lateral opening to the lumen along the length of the second elongated member, wherein the at least one first elongated member is positioned within the lumen of the second elongated member such that the distal tip of the at least one first elongated member extends through the distal tip opening of the second elongated member, and wherein the distal tip of the at least one first member forms a sealing engagement with the second elongated member between the distal tip opening and the at least one lateral opening of the second elongated member.

[0010] In one embodiment, the at least one first elongated member is positioned within the lumen of the second elongated member, and is sized such that a flow path extends from the at least one side opening of the second elongated member to the proximal opening.

[0011] In one embodiment, the device further comprises a third elongated member having a lumen extending between a proximal opening and a distal tip opening, wherein the third elongated member is sized to fit within the lumen of at least one first elongated member.

[0012] In one embodiment, the sealing engagement is formed by a gasket, a spacer, or a thread. In one embodiment, the distal tip opening of the at least one first elongated member is located at a distance of about 1 mm to 1000 mm from the at least one side opening of the second elongated member. In one embodiment, each of the at least one side openings extends along the second elongated member over a length of 5 to 25 mm.

[0013] In one embodiment, the at least one first elongated member is a plurality of members sharing a single proximal opening. In one embodiment, the at least one first elongated member is a plurality of members sharing a single distal tip opening. In one embodiment, the proximal openings of the at least one first elongated member and the proximal opening of the at least one second elongated member are each fluidly connectable to a solution reservoir. In one embodiment, the solution reservoir contains a composition selected from a cell recruitment composition, a pain relief composition, and a combination thereof.

[0014] In one embodiment, the cell recruitment composition includes an integrin family modulator, e.g., the VLA-4 molecule inhibitor firategast, UNII-OJY3SK9H5F, and BIO5192; a CXCL12 / CXCR4 interaction modulator, e.g., AMD3100; a CXCR7 molecule modulator, a CXCL12 analog; a neuronal / stem cell interaction modulator, such as through dopamine regulation or inhibition of neuronal axon firing; an adhesion molecule modulator; an integrin; a G protein-coupled receptor; an S1P-1 agonist; an endocrine target; and a prelixafor. This includes modulators of molecules in molecular pathways involved in cell adhesion or cell recruitment, selected from the group consisting of granulocyte colony-stimulating factor (G-CSF), pegylated and glycosylated versions of G-CSF, granulocyte-macrophage colony-stimulating factor (GM-CSF), macrophage colony-stimulating factor (M-CSF), tyrosine kinase 3 (FLT-3), ancestim, stem cell factors, cytokines (including interleukin-1, interleukin-3, interleukin-6, interleukin-7, interleukin-11, and interleukin-12), metalloproteinases, serine proteases, cysteine ​​proteases, peptidases, chemokines, multiple chemotherapies, such as cyclophosphamide, and combinations thereof.

[0015] In one embodiment, the pain-relieving composition is selected from the group consisting of lidocaine, prilocaine, tetracaine, benzocaine, procaine, mepivacaine, bupivacaine, etidocaine, tropacocaine, pipelocaine, stobaine, cyclomethylcaine, paretoxycaine, diclonin, pharicaine, pramoxin, amoranon, phenacaine, diperodone, dibucaine, and combinations thereof.

[0016] In another aspect, the present invention relates to a method of tissue aspiration, comprising the steps of: providing a suction device having at least one first elongated member having a lumen extending between a proximal and distal opening; at least one second elongated member having a lumen extending between a proximal and distal opening; inserting the suction device into tissue such that the distal openings of the at least one first elongated member and the distal opening of the at least one second elongated member are positioned near a biopsy site; administering at least one solution to a first region of the biopsy site through the at least one first elongated member; and withdrawing at least one aspirated material through the at least one second elongated member from a second region of the biopsy site adjacent to the first region.

[0017] In one embodiment, the insertion step is supplemented by administering at least one pain-relieving solution through the lumen of the at least one first elongated member. In one embodiment, the tissue is bone marrow tissue. In one embodiment, the tissue is adipose tissue.

[0018] In one embodiment, the above-mentioned at least one solution comprises an analgesic selected from the group consisting of lidocaine, prilocaine, tetracaine, benzocaine, procaine, mepivacaine, bupivacaine, etidocaine, tropacocaine, pipelocaine, stobaine, cyclomethylcaine, paretoxycaine, diclonin, pharicaine, pramoxin, amoranon, phenacaine, diperodone, dibucaine, and combinations thereof.

[0019] In one embodiment, the above at least one solution is an integrin family modulator, e.g., VLA-4 molecule inhibitor filategrast, UNII-OJY3SK9H5F, and BIO5192, a CXCL12 / CXCR4 interaction modulator, e.g., AMD3100, a CXCR7 molecule modulator, a CXCL12 analog, a modulator of neuronal / stem cell interactions such as by dopamine modulation or inhibition of neuronal axon firing, an adhesion molecule modulator, an integrin, a G protein-coupled receptor, an S1P-1 agonist, an endocrine target, a prelixafor, a granulocyte colony-stimulating factor (G-CSF), pegylated and glycosylated versions of G-CSF, The cell recruitment composition comprises a modulator of molecules in molecular pathways involved in cell adhesion or cell recruitment, selected from the group consisting of granulocyte-macrophage colony-stimulating factor (GM-CSF), macrophage colony-stimulating factor (M-CSF), tyrosine kinase 3 (FLT-3), ancestim, stem cell factors, cytokines (including interleukin-1, interleukin-3, interleukin-6, interleukin-7, interleukin-11, and interleukin-12), metalloproteinases, serine proteases, cysteine ​​proteases, peptidases, chemokines, combination chemotherapeutic agents such as cyclophosphamide, and combinations thereof.

[0020] In one embodiment, the first region and the second region are separated by a distance between 1 mm and 1000 mm. In one embodiment, the administration step and the withdrawal step are performed simultaneously. In one embodiment, the administration step includes the sequential administration of a CXCR4 inhibitor, VLA-4 or metalloproteinase or ISP-1 agonist, a compound that modulates neural stem cell regulation, and a molecule that impairs cell adhesion. In one embodiment, the administration step is performed first, and the withdrawal step is performed after a delay of 30 seconds to 120 minutes.

[0021] In one embodiment, the aspirate comprises one or more cells selected from the group consisting of hematopoietic stem cells, mesenchymal stem cells, epithelial stem cells, stromal cells, glandular cells, nerve cells, adipocytes, germ cells, and combinations thereof. In one embodiment, less than 10% of the one or more cells are blood cells.

[0022] In one embodiment, the average pressure change in the first region and the second region is between 15 and 50 mmHg.

[0023] In one embodiment, the method further comprises collecting a tissue sample within the lumen of at least one of the first or second elongated members.

[0024] In another aspect, the present invention provides a method of tissue aspiration, comprising providing a suction device having at least one elongated member having a lumen extending between a proximal opening and a distal opening, inserting the suction device into the tissue such that the distal opening of the at least one elongated member is positioned near the biopsy site, administering at least one cell mobilizing composition to the biopsy site through the at least one elongated member, and withdrawing at least one aspirate through the at least one elongated member.

[0025] In one embodiment, the insertion step is supplemented by administering at least one pain relief solution through the at least one elongated member. In one embodiment, the tissue is bone marrow tissue. In one embodiment, the tissue is adipose tissue.

[0026] In one embodiment, the at least one cell mobilizing composition comprises a modulator of the integrin family, such as the VLA-4 molecule inhibitor fingolimod, UNII-OJY3SK9H5F, and BIO5192, a modulator of the CXCL12 / CXCR4 interaction, such as AMD3100, a modulator of the CXCR7 molecule, a CXCL12 analog, a modulator of the nerve / stem cell interaction, such as via dopamine modulation or inhibition of axonal firing, a modulator of adhesion molecules, integrin, G protein-coupled receptor, S1P-1 agonist, endocrine target, prerixafor, granulocyte colony-stimulating factor (G-CSF), pegylated and glycosylated versions of G-CSF, granulocyte macrophage colony-stimulating factor (GM-CSF), macrophage colony-stimulating factor (M-CSF), tyrosine kinase 3 (FLT-3), anthestim, stem cell factor, cytokines (including interleukin-1, interleukin-3, interleukin-6, interleukin-7, interleukin-11, and interleukin-12), metalloproteinase, serine protease, cysteine protease, peptidase, chemokine, combination chemotherapy agents, such as cyclophosphamide, and combinations thereof, and comprises a modulator of a molecule within a molecular pathway involved in cell adhesion or cell mobilization selected from the group consisting of these substances.

[0027] In one embodiment, the at least one cell mobilizing composition further comprises an analgesic selected from the group consisting of lidocaine, prilocaine, tetracaine, benzocaine, procaine, mepivacaine, bupivacaine, etidocaine, tropacocaine, pipocaine, stobaine, cyclomethycaine, parethoxycaine, dyclonine, faricaine, pramoxine, amolanone, phenacaine, diperodon, dibucaine, and combinations thereof.

[0028] In one embodiment, the administration step includes the sequential administration of a CXCR4 inhibitor, a VLA-4 or metalloproteinase or ISP-1 agonist, a compound that modulates neural stem cell regulation, and a molecule that impairs cell adhesion. In one embodiment, the administration step is performed first, and the withdrawal step is performed after a delay of 30 seconds to 120 minutes.

[0029] In one embodiment, the aspirated material contains one or more cells selected from the group consisting of hematopoietic stem cells, mesenchymal stem cells, epithelial stem cells, stromal cells, glandular cells, nerve cells, adipocytes, germ cells, and combinations thereof. In one embodiment, less than 10% of the one or more cells are blood cells.

[0030] The following detailed description of embodiments of the present invention will be better understood in conjunction with the accompanying drawings. However, it should be understood that the present invention is not limited to the exact arrangement and means of the embodiments shown in the drawings. [Brief explanation of the drawing]

[0031] [Figure 1] Figure 1 shows the individual components of an exemplary suction device, including a cannula, trocar, and stylet. [Figure 2] Figure 2 shows an exemplary suction device configured to be drivable within the target suction site. [Figure 3] Figure 3 shows a cross-sectional view of the handle portion of an exemplary suction device. [Figure 4] Figure 4 shows cross-sectional views of the handle portion (left) and distal end (right) of an exemplary suction device. [Figure 5] Figure 5 shows an exemplary suction device having a lumen that can rotate independently of the access port while maintaining fluid connectivity. [Figure 6] Figure 6 shows a syringe fitted to the handle portion of an exemplary suction device, for example, in a perforation configuration. [Figure 7]Figures 7A and 7B illustrate the injection process during puncture using an exemplary suction device. In Figure 7A, the suction device is positioned near the suction site, and the fitted syringe is prepared with the injection solution. In Figure 7B, the solution is injected at the distal tip of the suction device. [Figure 8] Figure 8 shows a syringe fitted to the handle portion of an exemplary suction device, for example, in a suction configuration. [Figure 9] Figure 9 is a diagram of an exemplary device of the present invention demonstrating suction and injection. [Figure 10] Figures 10A and 10B illustrate the aspiration process using an exemplary aspiration device. Figure 10A shows the aspiration device inserted into the bone marrow, with a fitted syringe prepared for aspiration. In Figure 10B, directional aspiration is achieved using a single side opening. [Figure 11] Figure 11 shows two syringes fitted to the handle portion of an exemplary suction device, such as in a simultaneous suction and injection configuration. [Figure 12] Figures 12A and 12B illustrate the extraction of a biopsy core using an exemplary suction device. Figure 12A shows the cannula portion used to remove the biopsy core. Figure 12B is a magnified view showing the retention of the biopsy core via the threaded tip. [Figure 13] Figure 13 shows an exemplary pressure-regulating syringe. [Figure 14] Figure 14 shows an exemplary suction device cannula. [Figure 15] Figures 15A and 15B show a cannula and puncture instrument that fit an exemplary suction device. Figure 15A shows a trocar. Figure 15B shows a drill bit. [Figure 16] Figure 16 shows a multi-lumen stylet that fits an exemplary suction device cannula. [Figure 17] Figure 17 shows a single lumen stylet fitted with a cannula of an exemplary suction device. [Figure 18]Figure 18 shows an asymmetrical luminal stylet fitted with a cannula of an exemplary suction device. [Figure 19] Figure 19 shows a directional lumen stylet fitted to a cannula of an exemplary suction device. [Figure 20] Figure 20 shows an exemplary method for obtaining cells. [Figure 21] Figure 21 is a bar graph showing the vacuum pressure in bone marrow in a control (left) versus an exemplary suction device of the present invention (right). The exemplary suction device of the present invention reduced the pressure by 97%. [Figure 22] Figure 22 shows pressure measurements during three consecutive control aspirations performed using a 10cc syringe and a standard end-hole needle. Notice the minimum peak at 600 mmHg (perfect vacuum is -760 mmHg and near the negative pressure at which blood cavitates if gas spontaneously escapes the solution, impairing the effect). [Figure 23] Figure 23 shows an experimental run using an exemplary suction device of the present invention. Note the different scales. It allows visualization of normal positive perfusion pressure (~10 mmHg) of bone, with small peaks and dips in the baseline representing the heartbeat transmitted to the bone. This is a single experimental suction with the exemplary suction device combined with an equalization syringe. Notice the almost perfect equalization of pressure during suction. [Figure 24] Figure 24 shows the microanatomical structure of bone marrow. The right image is an electron micrograph of a bone marrow vascular cast, where the tubular structures are sinusoidal capillaries and venules filled with small amounts of blood. The blue stars represent the spaces in which stem cells are anchored to the parenchyma. When the needle is placed in the bone, it enters these vascular spaces. When suction is applied, blood, not the anchored stem cells, flows from these tubes into the needle. This is the reason for blood contamination, and why current bone marrow aspiration extracts only blood and not true stem cell harvesting. [Figure 25]Figure 25 shows a diagram of one molecular pathway demonstrating stem cell mobilization using enhancement by intraosseous drug administration, which achieves much higher initial peak concentrations at the site of action than would be possible with other administration routes. Drug administration allows cells to move out of their niches and into these vascular spaces, enabling harvesting of cells in greater numbers. [Figure 26] Figure 26 shows a 3D volumetric CT scan of a pig model in which an exemplary suction device was inserted into the bone and a radiopaque contrast agent was injected into the inlet, showing that the agent diffused over a large portion of the bone. [Figure 27] Figures 27A and 27B show the results of experiments comparing aspiration performance using the aspiration device of the present invention, the aspiration device of the present invention with a pharmacological recruiter, and the control method with a pharmacological recruiter. Figure 27A shows the colony-forming units (CFU) per 1 mL of aspirated material, which are calculated to account for differences in the number of cells in the aspirated material. Stem cells form colonies through replication and differentiation, while non-stem cells do not. Therefore, the number of colonies is a recognized measure of stem cell acquisition. When the cell number is controlled, both the pressure-relieving mechanism and the pharmacological recruiting mechanism result in a much higher number of colonies per leukocyte than the control, consistent with a reduction in blood contamination. When the stem cell recruiter is administered via the control device and aspiration is performed through the same route as drug administration, the cell number is significantly lower than even the control, demonstrating the importance of the device in the mechanism for drug delivery and collection. Figure 27B shows the results of Figure 27A normalized for cell number. When total colony count is measured rather than controlling for white blood cell count, the importance of pressure relief mechanisms and drug recruitment methods becomes clear because these methods yield a much higher total white blood cell count yield. Both device methods alone and those combined with pharmacological recruitment methods demonstrate significantly higher stem cell yields. [Modes for carrying out the invention]

[0032] This invention provides an improved device for biopsy, aspiration, and stem cell acquisition, and a method for using the same. The device balances aspiration and simultaneous injection to manage pressure changes at the biopsy site. The invention can be adapted to any biopsy, aspiration, or cell harvesting procedure, including adipose tissue aspiration and bone marrow aspiration (BMA). In particular, the invention increases cell mobilization, such as limiting patient pain, preventing blood contamination, and improving stem cell yield through intraosseous (IO) pharmacological mobilization of stem cells during BMA procedures, and improving stem cell yield using pharmacological mobilization of stem cells from adipose tissue. Pharmacological mobilization of cells allows for the harvesting of cells from biopsies many times larger than with existing methods.

[0033] definition The drawings and description of this invention are simplified to illustrate elements relevant to a clear understanding of the invention, but for the sake of clarity, many other elements commonly found in the art have been omitted. Those skilled in the art may recognize that other elements and / or processes are desirable and / or necessary for carrying out the invention. However, since such elements and processes are well known in the art and would not facilitate a better understanding of the invention, no description of such elements and processes is provided herein. The disclosure herein covers all such variations and modifications to such elements and methods known to those skilled in the art.

[0034] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as those generally understood by those skilled in the art to which the present invention pertains. Any methods and materials similar or equivalent to those described herein may be used in carrying out or testing the present invention, but only exemplary methods and materials are described.

[0035] As used herein, each of the following terms has the meaning associated with it in this section.

[0036] The articles "a" and "an" are used herein to refer to one or more (i.e., at least one) grammatical objects of the article. For example, "one element" means one or more elements.

[0037] As used herein, "approximately" when referring to measurable values ​​such as quantity or duration means that it includes variations of ±20%, ±10%, ±5%, ±1%, and ±0.1% from the specified value, because such variations are appropriate.

[0038] As used herein, "anesthetic" refers to a drug that causes reversible loss of sensation in a particular area of ​​a subject's body. An example of an anesthetic is lidocaine.

[0039] As used herein, “aspicular sample” refers to a liquid sample of bone marrow that does not contain “spicules.” Spicules are particles of parenchyma tissue containing stem cells used for analysis; without a spicule evaluation, the results are very limited and the stem cell yield is low.

[0040] As used herein, “bone access needle” refers to a device used to access the medullary cavity through the hard cortex of bone.

[0041] As used herein, “distal” refers to the lower end of the device away from the mounting point or origin. In the disclosed embodiments, distal refers to the end furthest from the healthcare professional when the device is introduced to the patient. As used herein, “proximal” refers to the nearest end of the device that is closer to the center of the body or mounting point. In the disclosed embodiments, proximal refers to the end closest to the healthcare professional when the device is placed on the patient.

[0042] As used herein, "dry tap" refers to the failure to obtain liquid bone marrow during bone marrow aspiration. This occurs due to needle misalignment or situations where venous blood within the bone is replaced by fibrous tissue or tumor cells. As used herein, "intraosseous injection" refers to the process of directly injecting a therapeutic agent into the bone marrow.

[0043] As used herein, "intramedullary space" refers to the space within the bone marrow cavity. As used herein, "lumen" refers to a tube, conduit, or cavity within a tubular structure. As used herein, "axially integrated" refers to a state in which a structure is integrated along its longitudinal axis.

[0044] As used herein, “channel” refers to a conduit, duct, or any kind of longitudinal hollow pathway used for transport in any longitudinal direction. For example, a channel may be used to deliver an anesthetic along the channel from a syringe to a target anatomical site, or a channel may be used to transport a tissue or cell sample along the channel from an anatomical site or lesion into a syringe.

[0045] As used herein, "injection" refers to the process of slowly introducing an element, such as a solution, into or onto a target. As used herein, “internal anatomical space” refers to any region and / or area located beneath the outer skin layer. Internal anatomical space may include cavities and / or cellular structures.

[0046] Throughout this disclosure, various aspects of the present invention may be presented in a range format. It should be understood that the range format description is merely for convenience and brevity and should not be interpreted as an inflexible limitation on the scope of the invention. Therefore, the range description should be considered to specifically disclose not only the individual numbers within that range but also all possible subranges. For example, a range description such as 1–6 should be considered to be a specifically disclosed subrange such as 1–3, 1–4, 1–5, 2–4, 2–6, 3–6, etc., as well as the individual numbers within that range, e.g., 1, 2, 2.7, 3, 4, 5, 5.3, 6, and any whole and partial increments between them. This applies regardless of the width of the range.

[0047] Suction device The present invention includes an aspiration device that can be adapted for bone marrow aspiration in cancer diagnosis, bone marrow harvesting, stem cell harvesting, intraosseous injection, trabeculotomy, bone biopsy, etc., for regenerative medicine or scientific purposes. Under the umbrella of regenerative medicine, the device of the present invention can also be used in orthopedic and spinal fixation devices. The device of the present invention can be used separately or combined within an orthopedic or spinal fixation system. For example, the device can be incorporated into a system for positioning pedicle screws to obtain bone marrow from the vertebrae during spinal fixation surgery. The device reduces patient pain, enhances procedural steps, increases bone marrow and stem cell sample yield, eliminates peripheral blood (hemodilation), and eliminates aspiration artifacts during core biopsy. The device regulates intraosseous pressure during these procedures, increasing yield and reducing pain, which can be an active mechanism (injection into bone) or a passive mechanism (negative pressure / suction from a device such as a syringe for harvesting bone marrow pulls fluid through a second channel, reducing or depressurizing the negative pressure from the suction). Its unique design also allows for the collection of bone particles using a "dry tap" setting for cancer analysis. The large lateral foramen design can reduce the rate of "non-bone fragment samples."

[0048] Referring to Figure 1, an exemplary suction device 100 is shown. The device 100 includes a cannula 110, a trocar 120, and a stylet 130. In some embodiments, the stylet 130 can be fitted into the trocar 120, and the trocar 120 can be fitted into the cannula 110, and the stylet 130, trocar 120, and cannula 110 may be concentric. In other embodiments, the stylet 130, trocar 120, and cannula 110 may be positioned adjacent to each other. For example, in some embodiments, the stylet 130, trocar 120, or cannula 110 may be glued or welded side by side. The device 100 may also include one or more brackets or slip fittings for releasably holding the stylet 130, trocar 120, or cannula 110 adjacent to each other. In other embodiments, the stylet 130, trocar 120, or cannula 110 may be used independently as separate components.

[0049] The cannula 110 is an elongated hollow tube having a cannula lumen 119 that extends throughout. The cannula 110 comprises a proximal handle 112, a distal open end 114, and at least one lateral opening 116 near the distal open end 114. The lateral opening 116 can be positioned at any suitable distance from the distal open end 114, such as between 1 mm and 1000 mm. A large single lateral opening 116 allows for rotational control of where aspiration is performed and enables a sampling area from a single opening that is up to 48 times or more larger than that of conventional end-hole devices. In some embodiments, the lateral opening 116 has an elongated shape to increase the likelihood of crossing islands of hematopoietic bone marrow containing stem cells rather than adipose bone marrow containing only fat. The lateral opening 116 can have any suitable length, such as between 5 and 25 mm. In some embodiments, the cannula 110 comprises two, three, four, five, or more lateral openings. The distal open end 114 may include one or more cutting teeth or flutes. The cannula 110 may have any suitable dimensions. For example, the cannula 110 may have a length and tube size comparable to bone marrow aspiration needles commonly used in the field, such as a length of 2 to 10 inches and a tube size of 8 to 19 gauge. In some embodiments, the cannula 110 may include markings placed at intervals along its outside to indicate the depth of insertion.

[0050] The trocar 120 is an elongated hollow tube having a lumen 129 that extends throughout its length. The trocar 120 comprises a proximal grip 122, a distal tapered end 124, and a threaded region 126 located near the distal tapered end 124. The trocar 120 is dimensioned to fit into the cannula lumen 119. Preferably, the outer diameter of the trocar 120 is smaller than the inner diameter of the cannula lumen 119 to allow fluid passage for purposes described elsewhere in this specification. As shown in Figure 4, the threaded region 126 can be engaged with the threaded region near the distal end of the cannula lumen 119.

[0051] The stylet 130 is an elongated hollow tube having a stylet lumen 138 that extends along its entire length. The stylet 130 comprises a proximal handle 132 and a distal open end 134. The stylet 130 is sized to fit into the lumen 129 of the trocar 120. In certain embodiments, the stylet 130 fits snugly into the lumen 129 of the trocar 120 so that there is minimal space between the stylet 130 and the trocar 120.

[0052] Referring here to Figures 2, 3, and 4, the proximal and distal structures of the assembly device 100 are shown. The proximal handle 112, proximal grip 122, and proximal handle 132 interlock to form a single larger handle. The distal tapered end 124 of the trocar 120 can be positioned to extend beyond the distal open end 114 of the cannula 110. The distal open end 134 of the stylet 130 can be positioned to extend beyond the distal tapered end 124 of the trocar 120. Thereafter, the combined distal ends form a puncture point suitable for drilling into bone.

[0053] Figure 3 shows a cross-sectional exploded view of the proximal handle 112, proximal grip 122, and proximal handle 132. This represents a triaxial assembly with a proximal open end 136 (connected to the opposite distal open end 134 by a stylet lumen 138) that receives a needle nested into a proximal open end 128 (connected to the opposite distal tapered end 124 by a trocar lumen 129) at the center of the cannula 110. The cross-sectional view shows the proximal open end 128 in the trocar 120 and the proximal open end 136 in the stylet 130. The cross-sectional view also shows the proximal lateral lumen 118 extending laterally from the proximal open end 117 of the cannula 110. The proximal lateral lumen 118 provides means of accessing the cannula lumen 119.

[0054] Figure 4 shows cross-sectional views of the proximal and distal structures of the assembled device 100. Notably, in the assembled proximal structure, the proximal grip 122 forms an airtight seal with the proximal opening end 117 so that the cannula lumen 119 extends fluidly from its distal end through the proximal lateral lumen 118. In the assembled distal structure, the threaded engagement in the threaded region 126 between the cannula 110 and the trocar 120 is visible. The gap between the inner diameter of the cannula lumen 119 and the outer diameter of the trocar 120 is also evident, and this gap allows for the movement of fluid through at least one lateral opening 116.

[0055] The proximal lumen 118 is shown in Figure 4 to be embedded in the proximal handle 112 and blocked by the handle 132, but those skilled in the art will understand that any implementation of the proximal lumen 118 is contemplated. For example, the proximal lumen 118 may extend through the proximal handle 132 and be accessible by an additional opening on the proximal handle 132.

[0056] In some embodiments, the proximal means for accessing the lumen of the device 100 is arranged such that the proximal handle of the device 100 can be operated or rotated independently of the lumen. In the first example, in some embodiments, the proximal lateral lumen 118 may extend from the side or below the proximal handle 112 as a separately rotatable structure so that the proximal handle 112, the proximal grip 122, and the proximal handle 132 can be rotated independently of the proximal lateral lumen 118 (not shown).

[0057] In the second example shown in Figure 5, the suction device 140 provides access points to both the cannula lumen 119 and the trocar lumen 129, which are rotatable independently of the cannula lumen 119 and the trocar lumen 129. The suction device comprises a proximal handle 142 connected to a fluid transfer drum 144. The fluid transfer drum 144 has a substantially cylindrical shape with a diameter and a longitudinal axis aligned vertically, and this cylindrical shape has a proximal portion and a distal portion. The fluid transfer drum 144 has a centrally located pocket at its distal end, which extends deep into the distal end of the fluid transfer drum 144. The fluid transfer drum 144 comprises at least one channel opening 146a located on the surface of the proximal portion and at least one channel opening 146b located on the surface of the distal portion.

[0058] At least one channel opening 146a opens into a centrally located channel aligned with the longitudinal axis of the fluid transfer drum 144, and the centrally located channel terminates in an opening located at the top of the distal end pocket of the fluid transfer drum 144. The opening located at the top of the distal end pocket of the fluid transfer drum 144 is connectable to the trocar 120 so that a fluid connection is made between the at least one channel opening 146a, the centrally located channel, and the trocar lumen 129.

[0059] At least one channel opening 146b opens into a pocket channel that terminates on the side of the pocket at the distal end of the fluid transfer drum 144. The pocket is connectable to a cannula 110 so that a fluid connection is made between at least one channel opening 146b, the pocket channel that terminates on the side of the pocket, and the cannula lumen 119.

[0060] The suction device 140 comprises an outer casing 148 having a substantially hollow cylindrical shape with an outer diameter, an inner diameter, and a thickness between the outer and inner diameters. The outer casing 148 has a longitudinal axis that is coaxial with and perpendicular to the longitudinal axis of the fluid transfer drum 144. The inner diameter of the outer casing 148 is dimensioned to be larger than the diameter of the fluid transfer drum 144, defining the space between the outer casing 148 and the fluid transfer drum 144.

[0061] The suction device 140 includes three O-rings 150a, 150b, and 150c, each O-ring having a thickness that ensures a secure fit within the space between the outer casing 148 and the fluid transfer drum 144. O-ring 150a is positioned proximal to at least one channel opening 146a, O-ring 150b is positioned between at least one channel opening 146a and at least one channel 146b, and O-ring 150c is positioned distal to at least one channel opening 146b. Thus, the space between the outer casing 148 and the fluid transfer drum 144 is divided into two fluidically isolated chambers surrounding the fluid transfer drum 144: a proximal liquid chamber 152a between O-rings 150a and 150b, and a distal liquid chamber 152b between O-rings 150b and 150c. The O-rings 150a, 150b, and 150c maintain a leak-roof fit while allowing the outer casing 148 and the fluid transfer drum 144 to rotate independently of each other.

[0062] The outer casing 148 further comprises a port 154a opening into the liquid chamber 152a over its thickness and a port 154b opening into the liquid chamber 152b over its thickness. A syringe 102 can be fitted into ports 154a and 154b, respectively. This establishes a liquid connection from the syringe 102 fitted into port 154a to the liquid chamber 152a, where the liquid can enter at least one channel opening 146a oriented in any direction during the rotation of the fluid transfer drum 144, flow through a centrally located channel, and into the trocar lumen 129. A liquid connection is also established from the syringe 102 fitted into port 154b to the liquid chamber 152b, where the liquid can enter at least one channel opening 146b oriented in any direction during the rotation of the fluid transfer drum 144, flow through a pocket channel into the pocket and the cannula lumen 119.

[0063] In Figure 6, syringe 102 is shown fitted into the proximal opening end 136 of stylet 130. In this way, the contents of syringe 102 can be inserted through the proximal opening end 136 so that they travel through the stylet lumen 138 and exit through the distal opening end 134. Syringe 102 can thereby deliver any suitable compound through stylet 130. In certain embodiments, syringe 102 can deliver an anesthetic such as lidocaine so that the anesthetic reduces pain immediately before the bone drilling procedure (Figures 7A and 7B), thereby ensuring direct delivery of the anesthetic to the bone penetration site.

[0064] In many aspiration procedures, total bone anesthesia is performed by intraosseous lidocaine injection, but the administration is painful due to the positive pressure changes caused by the injection. Referring here to Figure 8, the assembled device 100 is shown with the stylet 130 removed as a means of administering the injectable agent painlessly. The stylet 130 may be removed after the assembled device 100 has been successfully perforated at the target site. With the stylet 130 removed, the proximal lumen 118 and the proximal opening end 128 are exposed. A syringe 102 or any other solution source can be fitted into the proximal lumen 118. In this way, the syringe 102 or the solution source can be injected through the proximal lumen 118, through the cannula lumen 119, and out through at least one lateral opening 116 near the distal end of the cannula 110 (Figures 10A, 10B). In this configuration, the proximal open end 128 passively assists in pressure equalization at the injection site by allowing the fluid discharged from the injection site to enter the distal tapered end 124, flow into the lumen 129 of the trocar 120, and pushing out the replaced air from the proximal open end 128. Furthermore, having a decompression port allows for fluid and drug infusion without the positive pressure levels that could cause pain, eliminating the need for lidocaine. Those skilled in the art will understand that a similar effect can be achieved by fitting a syringe 102 or solution source to the proximal open end 128. In this process, the injection can be performed through the proximal open end 128, through the lumen 129 of the trocar 120, and outward from the distal tapered end 124, and the proximal lateral lumen 118 passively assists in pressure equalization at the injection site by allowing the fluid discharged from the injection site to enter at least one lateral opening 116, flow into the cannula lumen 119, and pushing out the replaced air from the proximal lateral lumen 118. Preferably, suction is not performed using the assembly device 100 shown in Figure 8. If the solution is suctioned from one proximal open end without supplying it to the opposing proximal open end, air or gas is introduced into the opposing proximal open ends, opposing lumens, opposing distal or lateral openings, and into the suction site from which a certain amount of suctioned material is removed.Air or gas can be drawn into the bloodstream, which can cause air or gas embolism and potentially cause injury to the patient.

[0065] Figure 9 shows the distal tip of device 100 having only a trocar 120 and a cannula 110 to illustrate the concept of pressure adjustment. As described elsewhere in this specification, commonly used bone marrow aspiration devices only provide the ability to aspirate, which causes pressure changes at the aspiration site, resulting in widespread pain, drawing blood into the sample and reducing the pressure to a vacuum. Device 100 can adjust the pressure changes by providing at least one additional lumen at the aspiration site. In Figure 9, the exemplary device 100 can administer a solution 10 from a solution source such as the aforementioned syringe 102 through the lumen 129 of the trocar 120. Device 100 can also withdraw the aspirated material 12 through the lumen 119 of the cannula 110. The distal open end of the cannula 110 is sealed by screw-engaging with the threaded region 126 of the trocar 120, which guides all the aspirated material 12 into the lumen 119 through the side opening 116. The flow of solution 10 and aspirated material 12 can be controlled independently, and as a result, administration and withdrawal can be performed simultaneously or intermittently as needed. Device 100 can switch between one or more solution sources to administer one or more solutions 10, such as a pharmacological solution from the first reservoir in the first stage and saline solution from the second reservoir in the second stage. Device 100 can also adjust the administration rate of solution 10 and / or the withdrawal rate of aspirated material 12.

[0066] In some embodiments, the syringe 102 may include a syringe fitted to the proximal lumen 118 and the proximal open end 128, as shown in Figure 11. In some embodiments, the syringe fitted to the proximal open end 128 may passively assist in pressure equalization by allowing the contents of the fitted syringe to enter the proximal open end 128, flow through the lumen 129 of the trocar 120, exit the distal tapered end 124, and replace the amount of aspirated material transferred. In other embodiments, the syringe fitted to the proximal open end 128 may actively assist in pressure equalization by actively pushing down the syringe fitted to the proximal open end 128, either manually or electrically. The syringe may dispense gas or liquid to assist in pressure equalization.

[0067] Referring here to Figures 12A and 12B, the trocar 120 can be removed, leaving the cannula 110 behind. With the cannula lumen 119 not obstructed, the core biopsy sample can be taken using the cannula 110. In certain embodiments, as described elsewhere in this specification, the distal open end 114 is provided with one or more cutting teeth or flutes that allow the cannula 110 to penetrate deeply into the biopsy site and capture the biopsy sample. A female threaded region near the distal open end 114 helps to hold the biopsy sample within the cannula lumen 119, so that the sample can be easily removed by withdrawing the cannula 110.

[0068] Referring here to Figure 13, an exemplary equalization syringe 200 is shown. The equalization syringe 200 includes a suction chamber 202 and a pressure regulating chamber 210. The suction chamber 202 is an airtight enclosure including a plunger 206 and a suction port 204 at its distal end. The pressure regulating chamber 203 is an airtight enclosure ending at an injection port 212 at its distal end. A fitting 208 fluidly connects the suction chamber 202 and the pressure regulating chamber 203 at its proximal end. In some embodiments, the pressure regulating chamber 203 includes one or more valves 214. One or more valves 214 can be used to introduce a solution into the pressure regulating chamber 203. The valves 214 can also be used to relieve excess pressure within the pressure regulating chamber 203. The valves 214 can be opened manually or passively in response to a given pressure level. In some embodiments, the pressure regulating chamber 203 can be subdivided into a plurality of interconnected spherical chambers 210, each having a valve 214, which can passively open to reduce the pressure within each sphere. For example, the size of the individual chambers 210 may affect the pressure at which the valves 214 open, such that larger chambers 210 discharge excess gas or liquid at lower pressures, and smaller chambers 210 discharge excess gas or liquid at higher pressures. In some embodiments, the suction port 204 and the pressure regulating port 212 are spaced apart so that the suction port 204 can be fitted into the proximal lumen 118 of the suction device 100, and the injection port 212 can be fitted into the proximal opening end 128 of the suction device 100.

[0069] In various embodiments, the equalization syringe 200 can automatically adjust the pressure at the suction site. When the plunger 206 is retracted proximal, the suction chamber 202 is filled with a certain amount of aspirated material entering from the suction port 204, and an equal amount of gas or fluid is discharged out from the proximal end of the suction chamber 202. The amount of gas or liquid exits the suction chamber 202 through the coupling 208 and enters the pressure regulating chamber 203, passing through the chamber 210 sequentially, so that the amount of gas or liquid displaces the contents of the pressure regulating chamber 203 and pushes the injected gas or liquid out of the injection port 212.

[0070] It should be understood that the equalization syringe 200 is not limited to the embodiment shown in Figure 13, as any mechanism for driving the contents of two adjacent chambers is contemplated. For example, in some embodiments, the suction chamber 202 and the pressure regulating chamber 203 may each have a plunger. The movement of each plunger can be mechanically linked, for example, by a lever arm or by displacement gas or liquid passing from one chamber to the other. The movement of each plunger can also be electronically linked so that a sensor can detect a value in one chamber and a controller can automatically reposition the plunger in the opposite chamber. Non-limiting examples of detectable values ​​include plunger position, amount of gas or liquid in the chamber, pressure in the chamber, etc.

[0071] In various embodiments, pressure control at the suction site can be performed passively or actively by having an attached device that combines negative pressure for recovery with positive pressure for adjustment. This combination can be at a ratio of -1:1 to maintain physiological pressure within the bone, or at any other ratio for a specific desired effect, e.g., -1:2 to "wash" the bone marrow away from the bone. Various embodiments may integrate valves to ensure that positive or negative pressure is not applied to the suction site via suction or injection unless certain parameters are met. For example, suction can be paused if the injection pressure is not at a desired ratio as the degree of vacuum to which it is applied. This may be a manual or automatic engagement valve system operating between the syringe and the suction and injection ports leading to the suction site.

[0072] The components of the suction device intended herein encompass multiple designs. Referring here to Figure 14, the distal end of a cannula 310 is shown. The cannula 310 includes an elongated hollow tube shape through which the cannula lumen 312 runs. The cannula 310 includes an open distal end 314 having at least one tapered rim 316. The cannula 310 further includes at least one side opening 318 near the open distal end 314.

[0073] Referring here to Figures 15A and 15B, various puncture instruments are shown. The trocar 320 has an elongated rod shape sized to fit into the cannula lumen 312 and terminates with a distal tapered end 322. In some embodiments, the trocar 320 includes a lumen 324 extending along its entire length. The lumen 324 can be used to directly administer drugs without requiring a separate stylet. In some embodiments, a drill bit 330 sized to fit into the cannula lumen 312 can be used as a trocar. In some embodiments, the drill bit 330 may further include a lumen extending along its entire length and can administer drugs similar to those in the lumen 324.

[0074] Referring here to Figure 16, an exemplary multi-lumen stylet 340 is shown. The multi-lumen stylet 340 comprises multiple lumens 342 and terminates at a stylet tip 344. The stylet tip 344 includes a substantially cylindrical shape having at least one side opening 348 and a closed end 346. The at least one side opening 348 may have the same length as at least one tapered edge 316 of the cannula 310. In some embodiments, the stylet tip 344 has a diameter dimensioned to fit snugly into the cannula lumen 312. In some embodiments, the stylet tip 344 includes one or more features to improve mating with the cannula lumen 312, such as a threaded area or a gasket. The multi-lubricated stylet 340 can be used to direct one or more injected gases or liquids through their respective lumens towards a suction site, such that the closed end 346 and at least one side opening 348 guide the injected gas or liquid laterally, resulting in suction closer to the side opening 318 of the cannula 310.

[0075] Referring here to Figure 17, an exemplary single-lumen stylet 350 is shown. The single-lumen stylet 350 comprises a single elongated tube having a lumen 352 running throughout and ending at a stylet tip 354. The single-lumen stylet 350 preferably has an outer diameter narrower than the inner diameter of the cannula lumen 312, resulting in a gap between the two to allow fluid movement. The stylet tip 354 includes a substantially cylindrical shape having at least one side opening 358 and a closed end 356. The at least one side opening 358 may have the same length as at least one tapered edge 316 of the cannula 310. In some embodiments, the stylet tip 354 has a diameter dimensionally sized to fit snugly within the cannula lumen 312. In some embodiments, the stylet tip 354 includes one or more features to enhance mating with the cannula lumen 312, such as a threaded area or a gasket. The single-lubricant stylet 350 can be used to direct the injected gas or liquid towards the suction site so that the closed end 356 and at least one side opening 358 guide the injected gas or liquid laterally, resulting in suction closer to the side opening 318 of the cannula 310.

[0076] Referring here to Figure 18, an exemplary asymmetric lumen stylet 360 is shown. The asymmetric lumen stylet 360 includes a single elongated conduit having a lumen 362 extending throughout and ending at a stylet tip 364. The conduit shape of the asymmetric lumen stylet 360 has a smaller cross-sectional area than the cannula lumen 312, resulting in a gap being formed between the two to allow fluid movement. The conduit can have any suitable cross-sectional shape, such as circular, elliptical, square, rectangular, or triangular. In some embodiments, the conduit has an arcuate cross-sectional shape, resulting in a cross-sectional shape having an arcuate side that can fit snugly against the inner surface of the cannula lumen 312 and at least one flat side facing away from the arcuate side. Thus, the asymmetric lumen stylet 360 is characterized by the ability to selectively close at least one side opening 318 of the cannula 310 using the arcuate side, thereby controlling the direction of fluid transport. The stylet tip 364 includes a substantially cylindrical shape having at least one side opening 368 and a closed end 366. The at least one side opening 368 may have the same length as at least one tapered edge 316 of the cannula 310. In some embodiments, the stylet tip 364 has a diameter that is dimensioned to fit snugly into the cannula lumen 312. In some embodiments, the stylet tip 364 includes one or more features to improve mating with the cannula lumen 312, such as a threaded area or a gasket. The asymmetrical lumen stylet 360 can be used to direct the injected gas or liquid towards the suction site so that the closed end 366 and at least one side opening 368 guide the injected gas or liquid laterally, resulting in suction closer to the side opening 318 of the cannula 310.

[0077] Referring here to Figure 19, an exemplary directional lumen stylet 370 is shown. The directional lumen stylet 370 includes a plurality of individual lumens 372 fixed to the stylet tip 374. Each lumen 372 extends beyond the stylet tip 374 and terminates at an open end 376 positioned at any appropriate distance from the side opening 318. The open end 376 can be oriented to face any direction. In some embodiments, the open end 376 can be actuated to change its direction or distance from the side opening 318. The open end 376 can also be actuated to macerate tissue at the suction site. In some embodiments, the directional lumen stylet 370 is at least partially composed of a shape-memory material such as nitinol, allowing each lumen 372 to take a predetermined shape after being inserted through the cannula lumen 312. The stylet tip 344 includes a substantially cylindrical shape having a diameter sized to fit snugly within the cannula lumen 312. In some embodiments, the stylet tip 374 includes one or more features, such as a threaded area or a gasket, to improve mating with the cannula lumen 312. A directional lumen stylet 370 can be used to direct one or more injected gases or liquids through each of its lumens 372 in any desired direction. In some embodiments, one or more lumens 372 can be used to transfer aspirated material from the suction site.

[0078] The various components of the present invention described above can be constructed using any suitable method known in the art. The manufacturing method may vary depending on the material used. For example, components substantially containing metal can be milled from a larger metal block or cast from molten metal. Similarly, components substantially containing plastic or polymer can be milled, cast, or injection molded from a larger block. Components substantially containing glass can be cut from a larger piece of glass. In some embodiments, the device can be manufactured using 3D printing or other additive manufacturing techniques commonly used in the art.

[0079] Suction method The present invention further includes an enhanced method of biopsy aspiration that increases cell yield and reduces pain. This method combines aspiration and solution administration and can be carried out using the aspiration device of the present invention as described elsewhere herein.

[0080] The method of the present invention is partly based on the surprising and unexpected finding that administering lidocaine using the novel device of the present invention in a BMA procedure increased stem cell yield by more than 200% compared to a control method. When lidocaine was administered using a standard BMA device, the stem cell yield decreased dramatically, as if the cells had been washed away from the site of lidocaine administration. However, when lidocaine was administered to a portion of the bone, and aspiration occurred remotely in a second portion of the bone, the stem cell yield increased significantly.

[0081] The method of the present invention is also based on the ability of the suction device to maintain relative physiological pressure within the bone during suction. This serves two purposes: 1) In other devices, the vacuum / pressure gradient extends from the syringe to the device, into the bone marrow, and into the blood, thereby aspirating less viscous blood and contaminating the aspirated material. On the other hand, this suction device passes the vacuum / pressure from the syringe to the device, into the bone marrow, and back to the syringe, removing mature blood cells from the aspirated material, and also, although an affirmative standard is not available due to venous contamination, 10 5 Doubling the CFU per individual cell shows a significant reduction in hemodilution. Furthermore, 2) since bone is highly sensitive to pressure changes that often cause pain, a decrease in vacuum reduces the pain experienced during suction.

[0082] Furthermore, administration of lidocaine has been shown to help reduce patient pain because it is a known anesthetic that blocks all nerve transmission, and it can create overall bone anesthesia during intraosseous injection.

[0083] Referring now to Figure 20, an exemplary method 600 is shown. Method 600 begins with step 602, which provides a suction device having at least one first elongated member having a lumen extending between a proximal and distal opening, and at least one second elongated member having a lumen extending between a proximal and distal opening. In step 604, the suction device is inserted into the tissue such that the distal opening of at least one first elongated member and the distal opening of at least one second elongated member are positioned near the biopsy site. In step 606, at least one solution is administered to a first region of the biopsy site through at least one first elongated member. In step 608, at least one aspirated material is withdrawn through at least one second elongated member from a second region adjacent to the first region of the biopsy site.

[0084] The insertion process may be carried out using any suitable means. For example, the suction device may be inserted using a trocar having a tapered distal end, or using a drill bit, as described elsewhere in this specification. The insertion process may be supplemented with the injection of an anesthetic to reduce pain, as shown in Figures 7A and 7B. The tissue may be any tissue containing the cells of interest, such as adipose tissue, bone marrow tissue, or the abdominal cavity. However, this method can also be used to recruit cells within any tissue of interest and is therefore applicable to all anatomical sites, such as the inner ear with auditory hair cells, the central nervous system with nerve cells, axonal cells, or supporting cells, the eye with retinal cells, or any of the various skeletal muscles, skin, teeth, heart, intestines, liver, or other organs and tissues. The cells of interest may be any suitable cells such as stem cells, stromal cells, glandular cells, nerve cells, adipocytes, germ cells, etc. The first and second regions may be directly adjacent or separated by a distance between 1 mm and 1000 mm.

[0085] In some embodiments, the administration and withdrawal processes are performed simultaneously. In this way, a certain amount of solution is continuously administered as a certain amount of aspirated material is withdrawn, so that the pressure within the tissue remains substantially constant. In one embodiment, the simultaneous action of solution administration and aspirated material withdrawal can be driven by actively pumping both media at their respective sites. In one embodiment, the simultaneous action of solution administration and aspirated material withdrawal is driven by the fluid pressure within the tissue. For example, active pumping of solution only into a first region increases the pressure within the tissue, where this increase in pressure drives the flow of aspirated material into a second region. In another example, active pumping (suction) of aspirated material only from the tissue from a second region decreases the pressure within the tissue, where this decrease in pressure drives the flow of solution into the first region. All of the above mechanisms demonstrate a reduction in pressure changes at the suction site in a pig model. Since pressure can be related to patient pain, it is preferable that the method results in an average pressure change of approximately 15–50 mmHg compared to a control value of approximately 400–700 mmHg.

[0086] In some embodiments, the withdrawal step follows the administration step after a delay. The delay allows the administered solution to penetrate the tissue and apply any number of therapeutic or cell-mobilizing effects to the tissue before the withdrawal of the aspirated material containing one or more cells. Multiple elongated members of the device of the present invention allow for the mitigation of pressure changes that may occur during injection and aspiration. The amount of solution administered can be less than or substantially equal to the expected amount of aspirated material, such as 1 mL to 15 mL of solution, depending on the required dose by the pharmacokinetics of the solution, and this can be based on approximately the patient's body weight. The delay time can be any appropriate time, such as a period between 30 seconds and 120 minutes.

[0087] In some embodiments, the administration and withdrawal steps can be carried out using at least one elongated member. For example, the method may include the steps of: providing an aspiration device having at least one elongated member having a lumen extending between a proximal and distal opening; inserting the aspiration device into the tissue such that the distal opening of at least one elongated member is positioned near the biopsy site; administering at least one cell mobilization composition to the biopsy site through at least one elongated member; and withdrawing at least one aspirated material through the same elongated member.

[0088] As described elsewhere in this specification, the administered solution can increase cell yield. The administered solution can act through one or more mechanisms, including, but not limited to, recruiting cells from their native environment, blocking inhibition of cell departure, reducing the adhesion of cells to their surrounding environment, and modulating the neural or cellular control that directs the stability, ingress, or egress of cells from their milieu. Compared to conventional procedures, the methods of the present invention can increase cell yield by at least 50%, at least 100%, at least 150%, at least 200%, at least 250%, at least 300%, or more. The solution may include any suitable composition that can increase cell yield by modulating the neural control of cell migration, by reducing adhesion to native tissue, or by producing a cell recruitment effect. Cells may be recruited into the surrounding extracellular environment / matrix, which may include the surrounding vascular space, and in the case of bone marrow, this may be into the surrounding capillary bed and sinusoidal capillaries. The solution may contain small molecules, peptides, polypeptides, nucleic acids, and carbohydrates. A non-limiting category of drugs that can be used includes currently known and undiscovered classes of proteins and receptors known to control cell migration out of tissue or cell retention within tissue (see Table 1). These may include α9β1 inhibition by combinations, e.g., BOP (antibodies or other small molecule selective inhibitors) and AMD3100. These encompass a broad category including: modulation of the integrin family such as the VLA-4 molecule inhibitors Philategrast, UNII-OJY3 SK9H5F, and BIO5192; modulation of CXCL12 / CXCR4 interactions such as the CXCR4 inhibitor Plerixafor; modulation of the CXCR7 molecule; and CXCL12 analogs.Modulation of neural / stem cell interactions, e.g., dopamine receptors (1-5 subtypes) and noradrenergic α and β receptors and all such receptors of catecholamines, their precursors and derivatives; modulators of their receptors or modulators of neurotransmitter uptake from synapses / sites of action; catecholamine degradation inhibitors, e.g., inhibitors of deamination by catechol-O-methyltransferase (COMT) or monoamine oxidase (MAO) enzymes; modulators of downstream cascades of catecholamine receptors such as adenylyl cyclase and alternative phosphoinositide 3-kinase (PI3K) / Akt pathway; and catecholamine receptors a1, a2, b1, b2, and b3. Gonists and blockers; catecholamine precursors such as L-phenylalanine, L-tyrosine, and L-DOPA; dopamine agonists, e.g., aripiprazole, phencyclidine, quimpyrole, salvinorin A, apomorphine, bromocriptine (Parlodel), cabergoline (Dostinex), siladopa, dihydroexidine, dinapsolin, doxanthrine, epicriptine, lislide, pergolide, pyribezil (Pronoran and Trivastal), pramipexole (Mirapex and Sifrol), propylnorapomorphine, quinagolide (Norprolac), ropinirole, rotigotine, roxindol, smanirole, phenoldopam selective for dopamine receptor D1, cocaine, amphetamine;Dopamine reuptake inhibitors, such as bupropion altropane (O-587), amhonelic acid (WIN 25978), amineptin (with a reasonable degree of selectivity for dopamine reuptake inhibition compared to norepinephrine), BTCP (GK-13), 3C-PEP (very potent and selective for dopamine transporters), DBL-583, difluoropine (O-620), GBR-12783, GBR-12935, GBR-13069, GBR-13098, GYKI-52895, and iomethopan (β-CIT). RTI-55), methylphenidate, ethylphenidate, modafinil, almodafinil, RTI-229, banoxerin (GBR-12909), haloperidol, chlorpromazine, eticlopride, pimozide, chlorpromazine, eticlopride; desipramine and other drugs that inhibit the reuptake of norepinephrine; DRD1, DRD2, DRD3, DRD4, DRD4 receptor agonists, and eticlopride Antagonists such as prido; nicotine; b2-adrenergic agonists such as clenbuterol; α9 integrin agonists; BOP, N-(benzene-sulfonyl)-L-prolyl-LO-(1-pyrrolidinylcarbonyl)tyrosine); VLA-4 antagonists, e.g., 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, e.g., Me6TREN; prolyl hydroxylase inhibitors, e.g., dimethyloxalylglycine (DMOG); chemokine GROβ; sulfated colomic acid; β-chemokine CCL15; Panax ginseng saponin (panax notoginseng saponins; VEGF; ALT-1188; P2RY14 agonists, e.g., MRS2690; UDP-glucose; γ-tocotrienol; TGFβ, TGF-β1, and Substance P; regulation of adhesion molecules such as VCAM-1; interaction with integrins such as VLA-4(α9β1);G protein-coupled receptors such as P2Y purine receptor-14; S1P-1 modulators including ACT-128800, SEW2871, GSK2018682, FTY720, MRS2690, and dopamine; various endocrine targets such as NOTCH protein (parathyroid hormone); granulocyte colony-stimulating factor (G-CSF) and analogues (filgrastim); PEGylated and glycosylated versions of G-CSF; granulocyte-macrophage colony-stimulating factor (GM-CSF); macrophage colony-stimulating factor (M-CSF); tyrosine kinase 3 (FLT-3); and ancestim; stem cell factors; AMD3100; TG-0054; KRP203; 4F-benzoyl-TN14003; POL6326; P2G, SDF-1β variant proteins; CTCE-0021; pepducins such as CS549 and ATI-2341; cytokines (e.g., interleukin-1, interleukin-3, interleukin-6, interleukin-7, interleukin-11, interleukin-12); metalloproteinases; serine proteases; cysteine ​​proteases; peptidases; chemokines, etc. Combination chemotherapy agents, such as cyclophosphamide, can also be administered intravenously, as is currently done.

[0089] [Table 1-1] [Table 1-2] [Table 1-3] [Table 1-4] [Table 1-5] [Table 1-6] [Table 1-7]

[0090] Various types of stem cells are needed for various applications. Mesenchymal stem cells are desirable for regenerative medicine purposes. Hematopoietic stem cells are desirable for bone marrow transplantation applications. Epithelial stem cells are desirable for vascular regeneration purposes. In certain embodiments, the solution comprises a composition selected for the extraction of a specific cell type, such as stem cells or stromal cells. This has been shown to be possible by the selective activation of one or more types of receptors, including activation / inhibition of the dopamine DRD2 receptor. In certain embodiments, the solution comprises a composition selected for the extraction of a specific stem cell type, such as hematopoietic stem cells, endothelial stem cells, and mesenchymal stem cells. For example, dopamine has been shown to increase hematopoietic stem cell recruitment while inhibiting mesenchymal stem cell recruitment. In certain embodiments, chemicals can be administered in various orders. For example, chemicals are designed to mobilize stem cells from their original niche (via a regulatory mechanism involving CXCR4, CXCL12, integrin inhibitors, or G-CSF, followed by proteases or protease inducers that commonly act on the cell's binding to the extracellular matrix, followed by drugs that act via the nervous system, nerves, or nerve receptors).

[0091] As described elsewhere in this specification, the administered solution may reduce pain. The solution may contain any suitable composition that can reduce the sensation of pain or mobilize stem cells. Non-limiting examples include one or more lidocaine, prilocaine, tetracaine, benzocaine, procaine, mepivacaine, bupivacaine, etidocaine, tropacocaine, piperacaine, stobaine, cyclomethylcaine, paretoxycaine, diclonin, pharikaine, pramoxin, amoranon, phenacaine, diperodon, dibucaine, and the like.

[0092] The various compositions described herein may be salts even if they are not indicated, and as will be well understood by those skilled in the art, the present invention is understood to include all salts and solvates of the indicated compositions, as well as unsalted and unsolvated forms. The various compositions described herein also encompass stereochemical forms, including any enantiomer or diastereomer forms of the indicated compositions. The enumeration of structures or names herein is intended to include all possible stereoisomers of the indicated compositions, such as crystalline or amorphous forms. Compositions may also include substantially pure compositions containing their particular stereochemical form, or compositions containing stereochemical mixtures in any proportion. The various compositions described herein also encompass analogues and derivatives that have similar structures to the indicated compositions but differ with respect to certain components or structural configurations, such as groups with linked atoms or longer or shorter linkers, or ring groups with different numbers of atoms.

[0093] The solutions of the present invention can be formulated using one or more pharmaceutically acceptable excipients or carriers. Useful pharmaceutically acceptable carriers include, but are not limited to, glycerol, water, saline solution, ethanol, and other pharmaceutically acceptable salt solutions such as phosphates and organic acid salts. Additional components may include, but are not limited to, one or more of the following: dispersants, inert diluents, binders, lubricants, preservatives, suspending agents, buffers, antioxidants, antibiotics, antifungals, and stabilizers.

[0094] In some embodiments, the method of the present invention can also harvest trabecular bone. For example, as the suction device advances to a fixed position, it cuts the bone fragments at its distal end, which has experimentally demonstrated scoop and relief angles, ideal for cutting bone with minimal force and minimal heat generation. These bone fragments are guided through a flute to a lateral opening of the device and scooped into the device lumen for harvesting. This has significant implications in regenerative medicine treatment, as both bone and bone marrow are often needed to assist in spinal fusion and other orthopedic procedures. Sometimes, the bone marrow is full of cells and liquid bone marrow cannot be obtained—a so-called "dry tap." Thus, the method for obtaining trabecular bone for analysis is useful even when bone marrow is unavailable. The device macerates the bone as it advances to the suction position, so the cells are "disaggregated," making them harvestable.

[0095] In some embodiments, the methods of the present invention can also induce a flushing action to facilitate the movement of substances away from a target site. For example, the devices and methods can be used to flush out calcium deposits in calcifying tendinitis and neoplastic calcification, or to non-invasively flush out joints for arthritis or infection. [Examples]

[0096] The present invention will be described in more detail with reference to the following experimental examples. These examples are provided solely for illustrative purposes and are not intended to limit the invention unless otherwise specified. Therefore, the present invention should not be construed as being limited in any way to the following examples, but rather as encompassing all variations that become apparent as a result of the teachings provided herein.

[0097] Without further explanation, those skilled in the art will likely be able to use the compounds of the present invention and implement the methods described in the claims by using the above description and the following exemplary examples. Therefore, the following examples specifically point to exemplary embodiments of the present invention and should not be construed as limiting the remainder of this disclosure.

[0098] Example 1: Intraosseous method for pharmacological mobilization of stem cells In a typical bone marrow aspiration (BMA) procedure, when stem cells are collected near the aspiration site, venous blood inevitably flows in from sinusoidal capillaries, contaminating the sample and limiting the number of stem cells that can be collected. Slow intraosseous injection of a drug allows it to diffuse throughout almost the entire bone, as bone is a large, interconnected venous space, much like a sponge. If the injection is too rapid, the drug is simply pushed into the venous system rather than the bone. By first pharmacologically overflowing the venous space of the bone, stem cells away from the aspiration site can be collected in sinusoidal blood, significantly improving yield beyond the limits of existing device configurations. Subcutaneous and intravenous (IV) pharmacological methods are too long to be useful for intraoperative collection required for regenerative medicine and cancer diagnosis. Intraosseous injection ensures peak concentration directly in the bone marrow, rapidly mobilizing stem cells that can be directly aspirationed. Preliminary data using the common and inexpensive drug lidocaine surprisingly and unexpectedly yielded a significantly increased yield of over 200% compared to the control (Figures 27A and 27B), demonstrating that lidocaine has a mobilization effect. The unique dual-lumen configuration of the novel BMA device allows for drug delivery at a site distant from the aspiration site, enabling drug diffusion to the aspiration site without washing away the stem cells to be aspirationed.Furthermore, lidocaine is effective for total bone anesthesia, which allows for painless suctioning (Manohar M et al., Veterinary Radiology & Ultrasound 17.4 (1976): 152-156; Tobias JD et al., Pediatric emergency care 6.2 (1990): 108-109; Waisman M et al., Journal of Trauma and Acute Care Surgery 39.6 (1995): 1153-1156; Replogle K et al., The Journal of the American Dental Association 130.5 (1999): 649-657; Chamberlain TM et al., General dentistry 48.3 (1999): 299-302; Joseph G et al., Journal of clinical anesthesia 20.6 (2008): 469-473; Ngo ASY et al., International journal of emergency medicine 2.3 (2009): 155-160; Philbeck TE et al., JEMS: a journal of emergency medical services 35.9 (2010): 58-62; Tobias JD et al., Anesthesia & Analgesia 110.2 (2010): 391-401; Sokov EL et al., Terapevticheskii arkhiv 85.4 (2012): 61-65). However, when lidocaine injection was performed with conventional devices, the yield was significantly lower than that of the control, indicating that stem cells at the injection site were washed away by the injection configuration of the conventional devices.

[0099] The colony-forming unit (CFU) assay is a recognized measure of the relative stem cell concentration in biomass ophthalmosin (BMA). Stem cells develop into colonies, while mature cells do not. A preliminary (N=4) BMA assay was performed in a live pig model, comparing a single-port cancer aspiration device with a novel dual-lumen BMA method. The novel dual-lumen device with pressure regulation provided higher CFU / mL than the control. The novel dual-lumen device with lidocaine injection (without pressure regulation) more than doubled the CFU / mL (Figure 27A).

[0100] Example 2: Eliminating sampling errors In cancer diagnosis, there are three main causes of sampling errors. The first cause is a "dry tap" (6.8% of aspirations) where bone marrow is not obtained. This can occur in normal patients, but it can also indicate serious disease, such as when the bone marrow is too densely packed with tumor cells to obtain fluid bone marrow. This can also occur after chemotherapy when all stem cells in the bone marrow have been removed, as well as in cases of fibrous bone marrow, such as in myelofibrosis. The second cause is non-bone fragment samples (20.6% of aspirations). This is the result of a small sampling area combined with the random distribution of bone fragment-containing hematopoietic bone marrow throughout the bone. It occurs when the sampling needle is placed in an area containing fatty bone marrow instead of red hematopoietic bone marrow. This bone marrow distribution is easily visible on MRI, but guiding each aspiration on MRI would be extremely cumbersome.

[0101] A third cause is hemodilution (27% of aspirated bone). Numerous small venous sinus-like capillaries flow into the bone cavity, which can be considered a large venous space. This venous space can sometimes even be mistaken for a vein in emergencies. This collection of blood flow is drawn into the aspirated vacuum. Early researchers have demonstrated that dilution of the bone marrow aspirate by peripheral blood and mature cells is unavoidable during bone marrow aspiration. Another study involving patients with blood disorders showed that 6–93% of nucleated cells were of blood origin, with the greatest mixing occurring in leukemia patients. When aspirating more than 2 mL, not aspirated as expected, much more bone marrow is not obtained; rather, venous blood preferentially flows into the needle for bone marrow due to its much lower resistance to flow from these venous channels. Numerous studies have demonstrated that only 1–2 mL of bone marrow can be reliably obtained from a single location, and that larger aspirated volumes from a single location reduce the yield. Another study found that the first 1.0 mL of bone marrow aspirated from a healthy donor contained 8% peripheral blood mononuclear cells (PBNCs), while subsequent aspirates for bone marrow harvesting contained 20% nucleated blood cells.

[0102] Experiments using 51Cr-labeled autologous erythrocytes and 125I-labeled albumin showed that approximately 97% of the hemoglobin in bone marrow aspirates originated from peripheral blood, regardless of whether it was at the start or end of aspiration, suggesting that bone marrow cells are aspirated within the blood volume. Importantly, depending on the number of trials required, up to 10–20 mL of sample may be needed for analysis, and more will be required if the patient is in a clinical trial where more bone marrow may be needed.

[0103] While this hemodilution is technique- and patient-dependent, it still has significant limitations despite optimized checklists and CT guidance. In particular, it confuses the ratio of mature to immature cells. Peripheral / venous blood from the bone marrow contains mature cells, while bone marrow itself contains both stem cells (immature) and mature cells. The ratio of these cells is the ultimate criterion for classifying myelodysplastic syndromes (MDS) and acute myeloid leukemia (AML), and therefore, dilution of this ratio by peripheral blood is a major factor reducing sensitivity in MDS and AML diagnosis.

[0104] In leukemia, blast count is used in diagnosis and prognosis / risk stratification, as well as as a diagnostic criterion for relapsed and residual disease. Hemodilution has been shown to negatively impact risk stratification in children with acute lymphoblastic leukemia. In a study evaluating minimal residual disease (MI) in treated acute myeloid leukemia, the blast percentage in hemodiluted samples changed to 83% compared to undiluted samples, and 4 / 9 (44%) of subjects were upgraded to a 0.1% cutoff for MI using undiluted samples. Peripheral blood contamination is important not only for searching for MRD but also at the time of diagnosis, when 25% of blasts are in the bone marrow and 0% are in the peripheral blood. Diluted samples show fewer than 25% blasts, which can lead to misdiagnosis.

[0105] In myelodysplastic syndrome (MDS), the blast threshold is elevated at any individual step of 5%, 10%, and 20%, and the possibility of category change depends on the proximity of the blast count to the threshold, in addition to the peripheral blood dilution of the sample. In one study of 66 MDS patients, attempting to suppress hemodilution resulted in a surprising reclassification of the disease in 33% of patients. Another study showed that peripheral blood dilution significantly limited flow cytometry in classifying MDS, with up to 26.8% of patients being reclassified after correcting for hemodilution. This correction is only applicable when bone marrow purity is 40% or higher.

[0106] As another effort to quantify the effects of hemodilution, a retrospective review was conducted of bone marrow aspiration and biopsy results from 355 MDS patients with at least one poor aspiration. The clinical significance of hemodilution aspiration was evaluated by comparing it to subsequent or repeated aspirations. Of 1250 aspirations, 470 (37%) were limited, of which 58% appeared clinically significant (22% of all aspirations in MDS), 4% resulted in missed important diagnoses, 7.7% required repeat biopsy, and 46% appeared clinically significant due to reduced confidence in blast counts or inadequate assessment for malformations.

[0107] Previous attempts to overcome sampling errors Dry Tap: Many authors suggest performing roll imprints against a dry tap background. This is when a core trefin biopsy is spread on a slide to simulate a bone marrow smear. However, these are highly technique-dependent, with effectiveness ranging from 10% to 60%. Another alternative method to complement dry tap is called "disaggregation," where an additional core biopsy is performed, and the entire volume of cells within that biopsy is isolated from the bone by either mechanical or enzymatic means, allowing for cytogenetics and flow cytometry as well as a substitute for a bone marrow smear. A 2005 study showed that 60% of samples yielded a good-quality aspirate-like analysis, and another 13% (75% total) yielded moderate but still beneficial quality. However, this requires patients to undergo additional or longer biopsies and is therefore not routine in clinical practice.

[0108] Non-fragmentary samples: An optimized approach using training and checklist interventions was employed to address the problem of non-fragmentary samples. In a 2013 prospective study, 18.5% of samples were non-fragmentary before the intervention, 20% were non-fragmentary after the intervention, and the non-diagnosis rate for non-fragmentary samples was 20%. Hematologists' unsupported confidence suggests that needle placement may be the cause of non-fragmentary samples. However, the use of CT scans at the iliac crest and complete placement did not improve the proportion of non-fragmentary samples in a subset analysis of retrospective examinations of sample quality.

[0109] Hemodilution: Attempts to overcome this long-standing problem of bone marrow aspiration date back to the 1960s, when lateral holes were added to the tip of the needle. Many still believe this increases the surface area for suction and therefore increases the yield of bone marrow rather than blood. However, since substances naturally flow where there is least resistance, blood still preferentially flows through the larger end holes of these devices or through lateral holes that may be connected to venous lakes in the bone marrow cavity. Randomized trials have shown that these lateral holes do not increase bone marrow yield, a fact confirmed by a 2011 study. Some data suggest that increasing the diameter of the needle core improves yield. However, no device adequately addresses the main problem. Devices with open end holes can only aspirate 1-2 mL from a single location in the bone without causing hemodilution. Such devices would have to be repositioned to different locations within the bone, imposing further pain and discomfort on the patient and therefore being impractical in clinical practice. Since current devices do not adequately address the problem of hemodilution, pathologists are trying to compensate for this fact using various means. This is not a standard treatment, but an attempt to manage dilution effects by using a combination of flow cytometry of both venous blood and bone marrow, which involves counting cells with a small number of cells and is only partially effective if the bone marrow sample is "sufficiently pure" (40-90% pure). The principal investigators suggest that these methods are impractical or not applicable in most cases.

[0110] Innovation to overcome sampling errors Dry Tap: As described above, "dissociation" of cells from trefin bone is a very effective means of obtaining aspiration-like analysis in the background of dry tap, but it is not used in clinical practice because there is no efficient way to deliver the sample and it requires an additional biopsy from the patient. Current needles compress the bone marrow tissue away from the needle as the needle advances through the bone to the aspiration position. The aspiration device of the present invention is designed to finely chop the bone through a serrated section as it advances to the aspiration position. The serrated section has a scoop angle and relief angle that has been shown to be ideal for cutting bone tissue with minimal force and minimal heat generation. These fragments are collected in a large aspiration chamber which itself has a short scoop angle and can be aspirated with the aspiration sample or removed in a stylet which has a ledge-like catchment that carries all solid fragments from the device when the stylet is removed (see Figures 16-18). These solid fragments can be placed in a relatively inexpensive machine for cell dissociation and subsequent aspiration-like analysis.

[0111] Non-bone fragment samples: As discussed, non-bone fragment samples are thought to be due to the placement of the aspiration needle in fatty bone marrow rather than hematopoietic bone marrow. There is a random distribution of hematopoietic bone marrow within the bone marrow, which can be sparsely scattered in many patients. This is one reason why the World Health Organization (WHO) requires a 2 cm bone core biopsy for proper diagnosis. To reduce this sampling artifact, the sampling length within the bone and the total volume of bone sampled must be increased to be equal to that of a bone core biopsy. Increasing the sampling length of the bone increases the likelihood that the needle will cross islands of hematopoietic bone marrow during aspiration to collect bone fragments. This increased sampling length can be achieved in the device of the present invention by aspirating through a single elongated lateral hole 10 mm in length, combined with the occlusion of the end hole during aspiration by the stylet remaining in place. This allows aspiration to occur around the stylet (see Figure 10B). Although other needles with lateral holes exist, the end hole, being the largest hole, is always the primary opening for aspiration. Furthermore, while other needles with lateral holes have them scattered around the needle, and therefore the user is uncertain about which hole actually harvested the bone marrow, the use of a single hole allows the user to control the rotation angle in which the hole faces. By blocking the end hole during aspiration, aspiration occurs across a defined length of lateral hole rather than at the end point. By holding the stylet, the needle can be moved in and out of the bone during or between separate aspirations. This allows aspiration to occur across a length of bone equivalent to that required by the WHO for the bone core.

[0112] Increasing the sampling area is achieved not only by the elongated holes, but the user can also rotate the needle and mix this increased surface area around the needle. By closing the end holes while holding the stylet during aspiration, exclusive aspiration from the lateral holes becomes possible. Thus, the user can control the bone aspiration area by controlling the rotation angle in which the aspiration chamber faces within the bone. Thus, the exemplary 10 mm lateral holes can be rotated six times for circumferential aspiration at each 10 mm depth position. Considering two depths within the same aspiration site, a 48-fold increase in the sampled area is explained.

[0113] Hemodilution: The main problem with hemodilution is that, with current methods that do not adjust negative pressure, only 1-2 mL can be aspirated from any location in the bone marrow before it becomes contaminated with peripheral blood. The clinical need is to be able to aspirate from multiple different regions within the bone marrow cavity within a single bone entry site. This, too, can be achieved, for example, by exclusive aspiration from a single lateral opening, occlusion of end openings, and the user controlling the rotation angle in the direction the aspiration opening faces. For example, six rotation positions at two different depths allow for more than 12 different aspiration positions. By limiting the amount aspirated per location to 1-2 mL, up to 24 mL of "pure bone marrow" can be obtained.

[0114] All patents, patent applications, and publications disclosed herein by reference are incorporated herein by reference in their entirety. While the present invention has been disclosed with reference to specific embodiments, it will be apparent that other embodiments and variations of the invention can be devised by those skilled in the art without departing from the true spirit and scope of the invention. The appended claims are intended to be construed as encompassing all such embodiments and equivalent variations.

Claims

[Claim 1] The invention described in this specification.

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