Patent Application
20210080358
The present invention relates generally to the field of medical and laboratory apparatus and methods of using the same. More specifically, the present invention relates to methods and apparatus for the extraction of tissue, cells, and other constituents from an enclosed body cavity such as a bone marrow cavity. Specifically, the present invention relates to methods and apparatus for increasing the efficiency of harvesting and processing mesenchymal stem cells, other biologic material, and physiological fluids for use in medical diagnostic, treatment, or research applications.
Mesenchymal stem cells, which for purposes of brevity will be referred to herein as (“MSC” or “MSC's”), are stromal or connective tissue cells which are found primarily in umbilical cord blood and bone marrow. Unlike hematopoietic stem cells (“HSC”) which contribute to the formation of red blood cells, MSC can differentiate into a variety of cell types such as bone, cartilage, muscle, and fat cells. While the cell differentiation phenomenon observed in MSC's is not fully understood, medical research has established that MSC and other constituents of bone marrow are remarkably capable of contributing to a patient's healing and tissue regeneration processes following injury or surgery and may be used therapeutically to treat numerous medical conditions.
MSC harvesting and processing cells and physiological fluid may be accomplished by several different conventional methods and apparatus. Exemplary methods and devices in current use involve multiple aspirations of bone marrow cells (BMC) from the patient's posterior iliac crest. Access to the internal cancellous bone, bone marrow and bone marrow blood residing inside the outer cortical bone layer of the ilium is achieved by creating an aperture in the cortical bone with a sharp instrument such as a trocar. An aspiration needle, for example, a Jamshidi bone marrow biopsy needle, is then inserted through the aperture into the interior trabecular compartments within the crest where the MSC are found in their highest concentrations. However, a significant problem associated with conventional methods for harvesting and processing cells and physiological fluid (including the aspiration of bone marrow from bone for producing stem cell concentrate) is that a less than desirable amount, type or quality of cells or physiological fluid may be harvested and subsequently processed while still maintaining cell viability. Substantial problems with respect to procedure efficiency, duration and desirable amounts, concentrations, purity, and viability of cells or biologic material remain unresolved.
Accordingly, a need exists for an improved system and method and apparatus for the harvesting and processing of cancellous bone, bone marrow and bone marrow blood for producing mesenchymal stem cell concentrate and other physiological fluids which addresses the combination of problems not solved by the prior art.
The stated problems and other needs in the art as apparent from the foregoing background may be addressed in accordance with the methods and apparatus of the present invention as set forth in various embodiments disclosed herein.
In one implementation of the present invention, a system for producing and maximizing the yield and viability of harvested tissue, by way of example, an enriched stem cell product, is disclosed which includes a bone core harvesting instrument, a modular receptacle or container adapted to receive the harvested bone core and any associated bone marrow, cells, and/or physiological fluid, a securing device or base member adapted to receive and secure the modular receptacle or container, a processing device adapted to process the harvested bone core in the modular receptacle or container, and a treatment syringe and needle.
According to an embodiment, the bone marrow harvesting instrument is a Jamshidi needle (also known as a trocar or stylet) movably deployed within an outer cannula having at least one externally-threaded end, the needle being adapted to create an aperture in the patient's cortical bone structure at a preselected location and to collect tissue, e.g., a bone core having a bony matrix, bone marrow, and bone marrow blood and deposit it into the modular receptacle.
In another embodiment, the processing device is configured to be secured by the securing device or base member and interface with the modular receptacle to process the harvested bone marrow, cells, and/or physiological fluid.
In yet another embodiment, the securing device or base member is adapted to be operatively secured to or mounted on a work surface such as a work or laboratory bench top.
In still another embodiment, the securing device or base member includes a manually powered apparatus configured to provide the mechanical power to process the bone core inside the modular receptacle.
In another embodiment, the securing device or base member includes an electrically or magnetically powered apparatus configured to provide electromechanical power to process the bone core inside the modular receptacle.
In an embodiment, the securing device or base member is configured to automate the processing of the bone core whereby the cells and tissue in the bone core are processed in a controlled manner pursuant to a preselected set of processing parameters.
In another embodiment, the modular receptacle or container is configured to receive and contain a carrier fluid.
In still another embodiment, the processing device includes a morselizing tool portion configured to morselize the bone core into smaller bone core pieces.
In yet another embodiment, the processing device includes a washing or rinsing portion configured to wash or rinse the bone core pieces whereby certain biologic material such as glycosaminoglycan (GAG) may be stripped from the bony matrix of the bone core pieces and put into solution with the carrier fluid.
In another embodiment, the washing portion is adapted to further break down the stripped biologic material into smaller predetermined sized portions.
In still another embodiment, the processing device may perform both the morselizing and the washing of the bone core with a single tool member.
In another embodiment, the single tool member may be actuated in response to preselected program processing parameters to perform the morselizing and the washing of the bone core.
In yet another embodiment, preselected program processing parameters include by way of example and not of limitation, morselizing, the single tooling portion may be located at a different location within the disposable module, operated at a different speed (whether rotationally or linearly reciprocated), or operated for a different duration, pulsed or continuous motion, and the like for the morselizing operation versus for the washing or rinsing operation.
In still another embodiment, the system of the present invention is structured and arranged for a single use, to be disposed of thereafter.
In another embodiment, the system of the present invention is structured and arranged to be sterilized for multiple use.
In an embodiment, a method for harvesting and processing a patient's mesenchymal stem cells is provided which includes inserting a Jamshidi-type needle, having a preselected diameter or size and a cannula extending circumferentially about and along the length thereof into the cortical bone at a preselected location on a patient's skeletal system forming an aperture therein; removing the stylet; advancing the cannula into the patient's bone marrow to obtain a cancellous bone plug or core; aspirating bone marrow material, blood, MSC and MSC-like cells through a cancellous bone plug positioned in the cannula to increase the harvested MSC cell count; and processing the harvested material in the system herein disclosed for producing and maximizing the yield and viability of an enriched stem cell product.
These and other features of the present invention will be apparent from the accompanying description of the invention, drawings, diagrams, and supplemental supporting materials provided herein.
The patent or application file contains at least one drawing executed in color. Copies of this patent or patent application publication with color drawing(s) will be provided by the Office upon request and payment of the necessary fee.
It should be noted that the present description is by way of illustration only, and that the concepts and examples presented herein are not limited to use or application with any single system or methodology. Hence, while the details of the system and methods described herein are for the convenience of illustration and explanation with respect to the exemplary embodiments, the principles disclosed may be applied to other types of tissue harvesting systems and methods without departing from the scope of the present invention.
Implementations of the present invention involve a system for processing a harvested tissue for preparing a biologic mixture. In particular, the system may include a collecting or harvesting tool for collecting tissue from a patient, a first module adapted to receive tissue from the harvesting tool and to process the tissue, a carrier fluid to receive and transport the tissue, a base module or member operably connected to the first or processing module, the base member being structured and arranged to be either hand-held or to be secured to a work surface or bench, and a collection and delivery apparatus or tool. The harvesting tool may include a tubular component configured to receive the tissue for collection and delivery into the disposable module. The harvesting tool may further include a plunger configured to force the tissue from the tubular component and into the base module. The various components, including both the first or processing module and the base member may be disposable or may be designed and constructed to be sterilized and reused for multiple patients and procedures.
Referring now to
The first module includes a handle or knob 22 adapted to be hand-operated or to be operatively connected to an electrically or magnetically powered apparatus configured to provide electromechanical power to process the harvested tissue such as bone core material and/or MSC following collection. As best viewed in
The base member 20 includes a slightly tapered cylindrically shaped body, preferably formed of a transparent material such a clear plastic or a polycarbonate material such as Lexan® to permit observation of the processing operation. The body has a wall portion 40 extending circumferentially about the axis A-A and extending intermediate upper and lower open ends 42, 44, the upper open end having a smaller diameter than the diameter of the lower open end, thus forming an isosceles trapezoidal-shaped cross sectional body configuration as shown in
A closure device, cap, or cover 48 is adapted to fit in removable sealing engagement into the open upper end 42 of the base member 20 in a manner analogous to a stopper or cork being urged into engagement with the end of a bottle or jar. In the embodiment of
Referring again to
The lower grinding plate 100 includes a plurality of radially outwardly extending fingers or vanes 112 formed integrally with or operatively connected to an upper surface 114 thereof. Similarly, as best shown in
Referring again to
A cylindrically shaped recess 140 is formed in the upper surface 134 extending circumferentially about axis A-A. The recess includes a bottom surface 142 and an inner surface 144 extending intermediate the bottom surface 142 of the recess and the upper surface 134 of the knob. An aperture 146 is formed concentrically about axis A-A extending through the bottom surface 132 of the knob and is adapted to rotatably and slidably receive the spindle 24, whereby the knob, spindle and upper morselizing or grinding plate may be controllably moved from an extended open configuration to a closed or depressed configuration. The spindle is releasably secured to the knob via a pin member 148 extending radially outwardly from wall portion 32 of the spindle and received in a notch or recess 150 formed in the aperture 146. A plug 152. A Luer connection 155 is formed on the first end 26 of the spindle and is adapted to provide a connection to a syringe or a filter and an interface for an automated an electrically or magnetically powered apparatus drive tool configured to provide electromechanical power to process the harvested tissue. A plug 158 having an elongate body 160 is releasably positioned in the coaxially extending cylindrical aperture 35 in the spindle and is adapted to prevent samples of harvested material from being trapped therein following injection or from being forced upwardly through the aperture during processing. The plug includes a grip or knob 162 operatively connected to a first end 164 of the plug, the grip having a female Luer connector 166 formed therein and adapted to fit in locking engagement with Luer connection 155 formed on the first end 26 of the spindle.
Referring now to
The morselizing/grinding procedure is performed with the upper grinding tool being rotated at a constant rotation speed of 180 RPM. Upon completion of processing, the fluid suspension containing GAG is collected and residual bone fragments are left in the receptacle. However, it is to be understood that the upper grinding tool feed rate and rotational speed may be selectively varied independently depending upon harvested material and desired end product characteristics and properties to optimize the process efficiency and quality of the resultant product.
Additional washing or rinsing steps may be performed as needed during the morselizing process to enhance the purity and output quantity of the product, which are illustrated in
Upon completion of the processing operation, the product is harvested as illustrated in
A hybrid morselizer plate design is 390 shown in
Yet another morselizer plate design is depicted in the embedment 400 of
Human-machine interface (HMI) may provide an auditory, tactile, textual, or graphical view of system conditions or operations and may be configured to allow an operator to monitor or control various aspects of the system. Alternatively, the HMI may be configured to report the status of various aspects of the system or process.
Motion control components selected and configured for accurate position/velocity/torque capabilities operating in either open or closed loop mode and may include servo systems, stepper systems, gearboxes, linear motion slides and actuators and drive couplings. A closed loop servo system may comprise feedback devices at the motor shaft to verify or adjust the resulting motion and may be configured to receive a command signal(s) for step and direction, CW/CCW, A/B Quad, internal indexing (for example, via discrete input and communication port), torque and velocity (for example, via analog input) and may include a control mode for torque, velocity and position.
The bench top configuration and/or other aspects of the system may comprise various power sources, programmable controllers, microprocessors, transmitters, encoders, gauges, meters, buttons, switches, indicators, interlocks or sensors having discrete or analog outputs configured to receive, send or provide data for automating various aspects of the system or process including rotary encoders for detecting position and speed; sensors for detecting proximity (inductive, magnetic, capacitive, ultrasonic), presence or distance (photoelectric), pressure, temperature, level, flow, turbidity, electrical conductivity, contrast, color, light transmission at one or move wavelengths, current and voltage; laser sensors, fiber optic sensors, fork sensors (or slot sensors); and limit switches for detecting states such as presence or end-of-travel limits.
The bench top embodiment and/or other aspects of the system may comprise various analyzers (to detect, quantify, profile, to characterize or measure the modular container constituents), metrological or cytometric modules, probes, components, reagents, assays, instruments, and systems. On-board cytometry may provide for the measurement or determination of the characteristics of cells for closed loop machine feedback or for operator display and may measure cell size, cell count, cell morphology (shape and structure), cell cycle phase, DNA content, RNA content, and the existence or absence of specific proteins on the cell surface or in the cytoplasm or otherwise characterize and count certain cells (for example red blood cells). For example, image cytometers, flow cytometers, cell sorters, time-lapse cytometers, assays, ELISA (enzyme-linked immunosorbent assay), real-time PCR systems may be incorporated in the system. In an embodiment, sampling plumbing may fluidically connect the chamber with one or more analyzers to selectably sample the container constituents before, during or after processing. In another aspect, the modular container may have an auxiliary draw port for manual sampling, and the sample may be delivered to a dry western blotting system for rapid analysis to monitor protein levels of, e.g., a-SMA and calponin. In particular aspects, a sample may be taken from the container through the plumbing and delivered to a real-time PCR module of the system in order to examine gene expression to determine if the shear stress caused by the cycle of the processing has markedly changed (e.g., increased) the mRNA levels of, e.g., CD31 and VE-cadherin. Such monitoring of gene expression may provide data in a control loop to control processing duration, speed and other parameters desirable for a particular patient, application, or other concern. The system may pause processing during sample analysis before either continuing processing or stopping processing depending on the data from the analysis.
The system is configured to collect, handle, process and deliver quickly, accurately, reproducibly, and reliably the biologic mixture from and back to a patient for an autologous point-of-care procedure. The disposable module may be configured, programmed, and controlled in a particular manner such that the washing cycle may impart a range of shear stresses on the stem cells or other constituents of the tissue or carrier fluid. It has been demonstrated that particular levels of shear stress will direct stem cell development along certain developmental pathways. The disposable module and processing system may be designed to selectively, repeatably and controllably impart a range of shear stresses on the stem cells to encourage development along numerous pathways so that the stem cells can be used in multiple applications autologously in the patient's body (Shear stress magnitude is critical in regulating the differentiation of mesenchymal stem cells even with endothelial growth medium-Biotechnol Lett. 2011 December; 33(12):2351-9. doi: 10.1007/s10529-011-0706-5. Epub 2011 Jul. 31.), which is incorporated herein by reference in its entirety, also see (Response of mesenchymal stem cells to shear stress in tissue-engineered vascular grafts-Acta Pharmacol Sin. 2009 May; 30(5): 530-536, published online 2009 May 5. doi: 10.1038/aps.2009.40), which is incorporated herein by reference in its entirety. The shear stress or an average shear stress may be in the range of 0.5 a to 1.5 Pa, 5 Pa to 100 Pa or 200 to 500 Pa. A maximum shear stress or a maximum average shear stress may be limited to about 0.5 Pa, 1 Pa, 1.5 Pa, 5 Pa, 15 Pa, 50 Pa, 75 Pa, 100 Pa, 200 Pa, 500 Pa, 700 Pa, 2500 Pa or 3000 Pa.
For example, shear stress and duration of exposure to the shear stress may be controlled by the grinding/washing tooling portion rotational speed, geometry or other parameters to maintain cell viability while being sufficient to detach certain material, e.g. GAG, from the bony matrix of the tissue and while reducing the particulate size to a desirable level. The maximum shear stress may be from about 0.5 Pa to about 1.5 Pa for a relatively long exposure duration; on the other hand, for short duration exposure, the maximum shear stress may be from about 700 Pa to about 2,500 Pa.
As an example, the chamber may be filled with about 1.75 milliliters of carrier fluid (e.g., normal saline) and about 100 milligrams of tissue. The washing tooling portion rotational speed may be run at about 3,000 RPM for about 100 seconds.
As another example, the chamber may be filled with about 1.75 milliliters of carrier fluid (e.g., normal saline) and about 100 milligrams of tissue. The washing tooling portion rotational speed may be run at about 15,300 RPM for about 10 seconds.
As noted above, the carrier fluid may comprise saline, autologous blood plasma, an anticoagulant solution, sodium citrate 4% w/v, platelet rich plasma (PRP) (leukocyte-rich PRP (L-PRP), leukocyte reduced PRP (P-PRP; leukocyte reduced or pure PRP), leukocyte platelet-rich fibrin or pure platelet-rich fibrin), growth factors, cytokines, ligands, hormones, neurotransmitters, RNA, plasmids, synovial fluid, exosomes or cerebrospinal fluid. The chamber may be purged or infiltrated with oxygen, nitric oxide, carbon monoxide or other gases or gas mixtures prior to carrier fluid and or tissue delivery or following delivery or during processing, and the material in the receptacle may be centrifuged prior to extraction.
Results:
Changes may be made in the above methods and systems without departing from the scope hereof. It should thus be noted that the matter contained in the above description and/or shown in the accompanying figures should be interpreted as illustrative and not in a limiting sense. The following claims are intended to cover all generic and specific features described herein, as well as all statements of the scope of the present methods and apparatus, which, as a matter of language, might be said to fall there between.
This application claims the benefit of U.S. Provisional Application No. 62/893,032, filed Aug. 28, 2019, the entire disclosure of which is incorporated herein by reference.
| Number | Date | Country | |
|---|---|---|---|
| 62893032 | Aug 2019 | US |
