Patent ID: 12247902

DETAILED DESCRIPTION OF THE INVENTION

The present invention enables the processing of excised tissue biopsy samples (TBSs) into a multitude of morcelized tissue particles (MTPs), for autologous wound healing. MTPs achieve effective healing by facilitating rapid cellular outgrowth from viable cells within a multitude of micrograft particles to radiate peripherally from and to bridge between the individual micrografts applied upon and covering an open wound.

The present invention embodies a sterile single-use disposable device which enables an effective and expedient means for a surgeon or medical practitioner to aseptically process an excised biopsy into an abundance of autologous micrograft particles. The process can be performed in an operating room or in a clinic at a patient's bedside. The procedure can be performed within a single patient visit procedure.

TBSs are processed into MTPs

for example, full thickness skin graft particles (FTSGPs), split thickness skin graft particles (“STSGPs”) and cartilage particles (“CPs”), in a fluid suspension for delivery to the autologous treatment area. This can be viewed as different stages, for example:
Stage 1 Harvesting

The first stage is to taking a tissue biopsy sample (TB S) from a patient for the purpose of processing the sample in accordance with the invention, into a morcelized form which can be applied to the area to be grafted (treated). A non-limiting example of a TBS may be an abdominal tissue biopsy sample, which can be taken and processed according to the invention. Other types of tissue biopsy samples may of course be taken, from different areas of the body and from different tissue types, including but not limited to cartilage graft biopsy samples, full thickness skin graft biopsy samples and split thickness skin graft biopsy samples. All such samples are included in the definition of TBS. These biopsy samples are relatively large compared to what they will be once they are morcelized and processed according to the invention.

Stage 2 Set-Up/Preparation

The TBS are added to a solution, such as saline solution, in the processing chamber of the system of the present invention. Desirably the solution is a sterile solution, e.g. sterile saline, or a solution buffered to a pH that is favorable to the tissue type and viability of the TBS.

Stage 3 Processing

The TBS are processed in the solution. The morcelization process results in the cutting of the TBS, using the inventive devices and processes, into viable morcelized tissue particles (MTPs) which are suspended in the solution. Examples of MTPs include FTSGPs, split thickness skin graft particles (STSGPs) and cartilage particles (CPs), among others. As stated throughout this specification, high viability of the MTPs is important to achieving a successful skin graft.

Stage 4 Separation/Dispensing

The MTPs in suspension are then filtered to remove the majority of the solution, which generally increases the viscosity of the suspension. While some solution remains, most has been removed and the resultant matrix of MTPs can be made to easily flow and spread on a wound or treatment area. Depending on the mesh size openings of the filter, which can be chosen to fit the desired graft application and tissue type, small MTPs will be removed along with the filtrate. This portion of smaller particles in the filtrate may be used for other applications such as injections for cosmetic use, or combined with additives such as anti-infectives or growth factors and used in the same application later on in the further treatment of the patient. The filtered portion of the MTPs are then conveyed or pushed (for example with a plunger or with suction) from the filter chamber into a dispenser (such as a removeably connected syringe), which is then used to apply or otherwise dispense the MTPs onto the area of treatment to form the new skin graft.

Stage 5 Application to the Treatment Site

Upon completing processing with the device, the MTPs are received into a fluidly coupled dispensing device, for example a dispensing syringe. The process can be completed with minimal user handling of the excised tissue. The matrix of MTPs in the dispenser is then ready for application to the treatment site. If the dispenser is a syringe or similar type dispenser, the matrix of processed MTPs can be readily applied by the physician or person administering the graft. The matrix of MTPs is intended to be easily spreadable and flowable, i.e. distributable, and the viscosity and density can be adjusted several ways, such as by determining the amount of solution to keep in the suspension during filtering or by later adjusting the density of MTPs distributed over the treatment site. Cellular outgrowth from the applied MTPs will adjacently form new graft tissue, ultimately bridging between and interconnecting individual MTPs to cover the area of treatment.

The device of the present invention is configured to be employed with a reusable, aseptically cleanable, operatable processor, used to drive the morcelizing mechanism within the device. Upon placing sterile saline and TBSs into the sterilized device, followed by activating the coupled operatable processor, the MTPs are aseptically processed within the device and then received into a fluidly coupled dispensing device, for example a syringe, ready for patient application. The viable tissue is maintained within the aseptic system, suspended within sterile fluid, (i.e. saline or buffered saline) throughout processing. Processing is completed within several minutes and free of user handling.

The device has features which optimize processing cell viability and convenience of tissue handling and transfer. Specifically, the aqueous processing allows for temperature, pH, and salinity control of the processing which would ideally be variable depending on the tissue, isotonic factors. The pH may be controlled with physiologic buffers. The variable blade speed allows for control of any potential baro-trauma caused by the formation of the vortex, which repeatedly moves the tissue suspended in the aqueous medium through the cutting devices.

The selection of ultra-sharp cutting blades is one important factor in ensuring that the tissue that is morcelized remains viable. The use of pH controlled aqueous medium, along with the ability to control the temperature of the medium during processing, as well as, the time, are also important factors in achieving morcelization with the exceptionally high degree of viability of the invention.

Mass-produced disposable razor blades and microtome blades are among the sharpest steel blades in the world. Razor type blades are typically martensitic stainless steel with a composition of chromium between 12 and 14.5% and a carbon content of approximately 0.6%. The high-volume linear process to produce such blades, starts with a roll formed strip of controlled thickness that is run as a ribbon through a continuous manufacturing process. The linear manufacturing process enables exceptionally tight and repeatable control of multiple sequenced automated processes including, for example, grinding multiple distinctly stepped beveled/faceted cutting blade edges on both sides; with cutting edges as thin as 30 nm for razor blades and 3 nm for microtome blades; with edges fortified with separate vacuum chamber applied hardened coatings (for example titanium+manmade diamond to harden edge), followed by, for example low friction polymer film for slipperier edge. Individual blades are progressively die-stamped in-line in a repeatable manner.

Blade sharpness, is absolutely necessary to minimize cell mastication. Use of ultra-sharp blades, passing through and between masses of live cells, and through interstitial spaces, best assures a narrow margin of violated cells along the cut line with the least amount of shear forces and crushing of cells. Slicing of tissue between two blade edges, passing at acute angles, enables stabilization of the tissue throughout each cut to achieve controlled atraumatic slicing of whole thickness skin tissue into morcelized particles.

Use of ultra-sharp blades, as achievable through automated processes, assures repeatability and expedites the morcelization process, enabling autologous whole thickness skin to be quickly converted into a new morcelized implantable tissue particles within minutes.

To maintain cell viability, best practice is to keep the harvested tissue wetted and then suspended in the pH controlled solution throughout handling (i.e. in saline, a buffer solution, or BioLife Solution®, or other cell nurturing/preservation solutions, etc.).

DETAILED DESCRIPTION OF DEVICES

One preferred embodiment for morcelizing tissue, such as full thickness skin grafts (FTSGs) and other tissues, supported in a fluid preferably sterile fluid, is shown generally inFIGS.1-4.

Processing device10includes a liquid-tight container12having an open upper end14which may be suitably enclosed by a cover16. The cover16has an inlet aperture18which allows for insertion of tissue into the fluid. In a preferred embodiment the container is generally cylindrical having a closed curved bottom20opposite open upper end14with and exit opening17therein. While the cover16and the container12may be made of various materials, in a preferred embodiment, the cover and container are formed of a suitable plastic such as polypropylene (PP), polyethylene (PE), polystyrene (PS), polyethylene terephthalate (PET), polyimide (PA), acrylonitrile butadiene (ABS), polyetheretherketone (PEEK) and polyurethane (PU). Combinations or co-polymers of these polymers may be used. Glass, ceramic or metal containers may also be used. The container12may be transparent to visualize the processing and quality of the fluid and tissue being processed within.

An additional removable or adjoined protective cover that is able to be manually opened and closed may be included so as to close off the cover opening during processing to best assure containment of fluid and cellular contents.

It is also contemplated that all components comprising the overall processing device (system), including the container and morcelizing mechanism, isolation device and applicator will be packaged and bulk sterilized, for single-patient use and disposable. The packaged devices may be irradiated with gamma or e-beam or ethylene-oxide (EtO). Alternatively, the processing device may be sterilizable and reusable. Some portions of the system may be reusable and other portions may be disposable.

Extending from bottom20, container12includes a generally elongate tubular conduit22in fluid communication with the interior13of container12through opening17. The conduit terminates in a container mount24at the lower end thereof. Extending outwardly and in fluid communication with conduit22is an outlet26which in the preferred embodiment shown inFIG.1extends at a right angle to conduit22. The description of the purpose of the conduit22, the container mount24and outlet26will described in further detail below.

Cover16is movably supported at the open upper end14of container12for movement along a central axis A. The upper end14of container12includes, for example, an outwardly directed key12awhich is seated in a slot16ain adjacent skirt16bcover16. The key12ais movable along axis A within the slot160to allow for the movement of the cover16with respect to the container12, while restricting cover16rotation about axis A.

The cover16further includes an inwardly formed downwardly extending generally tubular stem28having an upper cup-shape cavity30covered by a cap31. The stem28accommodates a mounting rod32having a threaded lower end34and an upper end36terminating in an enlarged head38. The head38is captively retained within the cavity30supported by a spring39, for example by one or more Belleville, dome, single or multiple wave type washers, captive between the lower end of cavity30and the enlarged head38. The spring or springs39may additionally be captively sandwiched between conventional type washers39b, the threaded lower end34of rod32is threaded into impeller108about axis A. A shoulder32b, adjacent to threaded end34of rod32, is supported against impeller108.

A disk shaped stationary cutting member102is supported upon the terminus of stem28on a perpendicular plain relative to axis A. The stationary cutting member102is constrained from rotating about axis A by, for example, mating pins28aor other keyed features in engagement between the stem28and the stationary cutting member102.

Now as best shown byFIGS.1-4andFIGS.9-10, the sub-assembled rod32, head38, impeller108, and blades112are together captured and configured to rotate about axis A relative to stem28. The spring or springs39, between head38and the lower end of cavity30, act to lift head38and thereby lift stem28and impeller108through stem38to continuously constrain rotating blade edges112ain compression against stationary cutting member102.

Referring particularly toFIGS.5-11B, the subassembly of rotating impeller108with associated rotating blades112, held compressively against stationary cutting member102, about axis A by means of a rod32with head38and lower end34and spring39through stem28are collectively referred to as a morcelizing mechanism.

As described in further detail below the drive shaft42is attached to an operatable processor44shown schematically inFIG.1by way of a drive engagement46at the lower end of drive shaft42. The processor44causes rotation of the drive shaft42and thereby rotation of the rotating blades112against the stationary blade edges122within the morcelizing mechanism40which causes morcelization of the tissue, or specifically FTSGs within in the container12. A suitable rotary shaft seals47and41provide a fluid seal between drive engagement46and drive shaft42.

The processor44is preferably a reusable device that is configured for ease of being aseptically cleaned following each use. The processor44is also configured to replaceably receive a processing device10in mechanical engagement in such manner that enables a user to attach, use, and remove the processing device using standard practices for interfacing surgical devices in a sterile field. The processor44may also be configured to receive a sterile drape to isolate surfaces not shrouded by a coupled processing device. The processor may also be a device that is entirely sterilizable and/or powered by compressed air that is easily available in the operation room setting.

As shown inFIG.2, and described in more detail below, rotation of the impeller108provides a continuous circulating flow (CF) of the fluid and contained tissue or specifically FTSGs about the interior of container12and through the morcelizing mechanism40so as to continually cut the FTSGs into progressively smaller particles. The morcelizing mechanism40is seated in fluid-tight relationship over exit opening17in the open bottom20of container12to maintain the FTSGs and fluid within the interior of container12throughout morcelization. A suitable seal19provides a fluid seal between morcelizing mechanism40and exit opening17.

As is shown inFIGS.3and4, the drive shaft42may be raised so as to unseat the morcelizing mechanism40from opening17in the open bottom20of container12. Upward movement of drive shaft42along axis A causes upward movement of the cover16with respect to the container12with the key12ariding within slot16a. This lifts morcelizing mechanism40off of its sealed position on the bottom20of container12thereby rendering accessible exit opening17for fluid flow.

Morcelizing Mechanism

Operative components of morcelizing mechanism40are shown in further detail with additional reference toFIGS.5-8.

Morcelizing mechanism40includes a base component100and a stationary cutting member102which are axially aligned over one another along axis A. Base component100is mounted to the drive shaft42with a depending mount104to provide for rotation. Above mount104is a flat circular plate106which is generally transverse. Plate106also serves as the seating surface in opening17of the bottom20of container12as is shown inFIGS.1-4.

The upper end of base component100serves as an impeller108having two or more impeller vanes110upwardly extending from plate106on diametrically opposed sides of axis A. The impeller vanes110are each curved along axis A in a complimentary fashion for purposes that will be described in further detail below. Each impeller vane110supports in facing relationship at the upper end a cutting blade112. As also will be described in further detail below, the cutting blades112at the upper ends of impeller vanes110are supported in juxtaposition with the stationary cutting member102. The blades112rotate with base component100with respect to stationary cutting member102.

In a preferred embodiment shown inFIGS.5-8, the stationary cutting member102has generally disc shaped body102a. The body102adefines spaced apart blade surfaces120arranged circumferentially. Each blade surface120includes a pair of converging blade edges122which converge at an apex122a. In between each of the blade surfaces, breaches124are defined. The breaches124are open spaces between the blade surface which permits passage of the TPs, such as FTSGs and other TPs, and fluid through body102aas the base component100rotates.

In one embodiment shown inFIGS.7and8, the stationary blade edges122are defined by longitudinal radially extending members123converging with an arc of the circle forming the outer edge of the disc shaped body102a.

In a more preferred embodiment shown inFIGS.5and6, the blade surfaces120are formed in a tear drop shape where the apex122aof the converging blade surfaces120converge near the circumference of the stationary cutting member102in a tapered curved surface. It has been found that this shape helps promote complete morcelization of the tissue and specifically TPs passed through.

The arrangement of the stationary cutting member102with respect to the impeller108is shown schematically inFIGS.9-12. A small clearance space (S) is provided between the lower edge of the stationary cutting member102and the upper end of impeller108such that the extending rotating cutting blades112are supported in juxtaposition against the lower edge of the disc shaped body102aof stationary cutting member102. This creates a shear plane (SP) at which the tissue is sheared and morcelized.

The stationary cutting member102is preferably stainless steel and CNC machined with precision ground sharp burr free stationary blade edges122. The bottom shearing plane (SP) surface must be flat and preferably 0.08 μm or better finish. The stainless steel material may generally be a corrosion resistant and hardened grade, for example 440C stainless steel, machined in annealed state and vacuum heat treated to 55-60 RC to achieve a hardened surface and durable sustainable cutting edges. The stationary cutting member102may alternatively be of other non-corrosive materials or may be of an alternative hardness and may be made by other precision process. The stationary cutting member102and juxtaposed rotating cutting blades112, in compressive engagement, may be of differing materials, such as, for example, plastic or ceramic, or of differing hardness, or have alternative treated, or applied surface finishes, to best avoid wear or galling conditions as opposing surfaces slide upon each other along a shear plane (SP). Additionally, the blade edges122of the stationary cutting member102and/or the blade edges112aof the rotating cutting blades112may be formed to be sharp or subsequently sharpened.

The rotating cutting blades112may be mounted at the upper end of the impeller108supported by a spring such as an elastomeric pad130which biases the edge of the cutting blade112against the lower edge of the disc shaped body102aof stationary cutting member102. It has been found that maintaining the cutting blade edges112ain physical contact against the stationary cutting member102, minimizes tearing and shredding of the tissue.

Referring toFIGS.5and6, the stationary cutting member102may have two or more breaches124radially arrayed about axis A, preferably as shown three, each with associated blade edges122. The rotating base component100may similarly have two or more cutting blades112radially arrayed about axis A, preferably as shown two. However, the quantity of rotating blades112best differ from the quantity of stationary blades122, so as to maximize cutting efficiency by minimizing otherwise cumulative cutting forces as would be compounded should multiple blades engage simultaneously.

The rotating cutting blades112are positioned at an acute cutting angle relative to the juxtaposed stationary blade edges122, such that tissue, when captured between the rotating blades112and juxtaposed stationary blades122will be cut with a slicing action.

FIG.12Ashows that cutting edges112aof the rotating blades112may be manufactured with a ground double beveled edge. Double bevel refers to beveled on both sides of the blade. Alternatively, as shown inFIGS.12B and12C, the cutting edges112acan be made sharper with a secondary distal honed double beveled edge which further maximizes the morcelization of the TFSGs. Honing refers to a more precise abrasive grinding or lapping process in which a relatively smaller amount of material is removed from the surface by means of a finer grit abrasive. The cutting blades112used in our functional proof-of-principle systems utilize preferably further sharpened blades which have a secondary honed double beveled edge, as well as an additional finely honed double beveled edge, for example three graduated sets of double beveled edges.

The rotating cutting blade112is best arranged at an acute angle relative to the lower surface of the stationary cutting member102, so that the tip of rotating blade cutting edge112apasses at an acute angle with respect to the stationary blade edges122of blade surfaces120.

As previously described above, springs39may be used to compressively preload the rotating blade edges112ato maintain contact upon stationary blade edges122throughout rotation.FIG.12Cshows that the cutting edge112aof the rotating blades may flexibly conform under preload against the shear plane (SP), particularly when the cutting blade112may be substantially stiff, for example approximately 0.010 inch thick. Additionally or alternatively, as shown inFIG.9, the cutting blade112, itself, may flexibly conform under preload against the shear plane (SP), particularly when the cutting blade112may be substantially flexible, for example approximately 0.003 to 0.006 inch thick.

FIG.12Dshows an embodiment where the blade edges112aof the rotating blades112may be formed to have a flat or planar extent112b. This planar extent112bis formed to be co-planar or co-extensive with the blade edges122of the stationary cutting member102at shear plane (SP).

While in a preferred embodiment, the shear plane (SP) is normal to the chamber impeller and blade rotation access. The shear plane (SP) may also take other direction with respect to the axis A. One example is shown inFIG.13Awhich shows a bushing200, a rotatory seal210, a stator220, a rotor240and a cutting chamber260. The blades242of rotor240pass in close proximity to the blades222of stator220which are stationary blades about a central axis A.

Also, in this embodiment, the rotor blades242have surfaces which are configured as integrally formed impeller vanes. The rotating edges may be generally co-extensive to the leading impeller vane edges. The rotating rotor blades and stationary stator blade edges should preferably remain in intimate physical contact to best achieve precise slicing. The blades may be machine honed for closely controlled minimum shear gap, preferably less than 30 micrometers. Positioning the rotating blade edges at an acute angle relative to the shear plane of stationary cutting blade edges facilitates a shear cut for the impinged tissue. Maintaining a spring assisted compressive engagement between the rotating blade edges and the shear plane of the opposing blade edges best assures that tissue will be precisely slice, rather than to slip between the converging blade edges.

Other techniques and arrangements for cutting the tissue at a shear plane may also be within the contemplation of one skilled in the art.

Tissue Morcelization

Having described the basic components of the process device10of the present invention, one preferred example of the morcelization of tissue into particles (MTPs) or specifically tissue particle (TPs) as defined herein will be described with respect to the Figures. The term “tissue particles” is also refereed to herein as TPs.

Initially, with reference toFIGS.1-4, FTSGs prepared as above described and in a fluid medium may be inserted into container12through inlet aperture18of cover16. A fill line F may be provided so to provide guidance as to the volume of tissue and fluid which may be placed in container12. Thereafter, the rotating mechanism connected to drive engagement46is activated so as to cause rotation of drive shaft42and morcelizing mechanism40.

Referring toFIG.2, such rotation causes circulating flow of the tissue in the fluid by establishing a vortex within container12. This vortex provides for continually moving the tissue through the morcelizing mechanism so as to fully morcelize the tissue contained therein. The circulating flow path as well as the vortex established is created by the configuration of the impeller vanes110of the impeller108.

Shown schematically inFIG.14, the impeller vanes110are constructed so that an upper or leading portion110aof the impeller vane110imparts an axial thrust upon the fluid and contained tissue (i.e., TBS) or specifically FTSGs, while a lower or terminal portion110bof the impeller vane110provides for radial thrust. The construction of the impeller vanes110aand110bprovide for continually moving the FTSGs throughout the container12. The impeller108causes fluid with contained FTSGPs to be driven through breaches124in the stationary cutting member102, passing between rotating blades112and stationary blades122, to then be deflected against the trough-like bottom20and side walls of container12to reverse the flow in the opposite direction, circulating through the outer peripheral volume (PV). The fluid flow then transitions into a vortex to mix and drive the tissue through the central volume (CV) to return again through the morcelizing mechanism40so that the FTSGs are continually and repeatedly cut and morcelized.

A person of ordinary skill in the art will be able to alternatively configure, for example, impeller vanes and/or internal container geometry and/or bottom20forms so as to enhance effective circulating flow of fluid with suspended tissue though-out the container12and through the morcelizing mechanism40. Internal flow characteristics may be enhanced, for example, by increasing or decreasing the pitch or otherwise reshaping the form of impeller vanes110; or by increasing or decreasing the pitch of a portion of a vane configured for axial thrust110arelative to the pitch of a portion of a vane configured for radial thrust110b; or to eliminate either of the axial thrusting vane surfaces or radial thrusting vane surfaces.

Referring toFIGS.3and4, once the TPs, such as FTSGs and other tissue particles as described herein, are fully morcelized, the drive shaft42may be raised, unseating the impeller108from its seated position in the container. The plate106is unseated from opening17establishing fluid communication with conduit22and outlet26. The morcelized tissue is discharged by a gravity driven drain through outlet26for use in a manner which will be described hereinbelow.

Upon completing the morcelization of FTSG or other tissues grafts into MTPs, the impeller108rotation may be stopped and the MTPs, as defined herein, having a specific gravity greater than water, will settle to the bottom20of container12. One skilled in the art will recognize that alternative methods may be used to manually withdraw the settled MTPs from the bottom20of container12within the processing device10. For example, the settled MTPs may be drawn into a conventional syringe in combination with an elongated cannulated tip (not shown) that can be inserted into the container12through the inlet aperture18. In such manner, the same syringe used to draw the MTPs from the processor10could then be used as an application device. In this manner of manually drawing out the MTPs through the inlet aperture18, the processing device10need not include a drain39or an outlet26.

Discharge of Morcelized FTSGPs

Discharging the morcelized Tissue Particles (MTPs) or for example specifically FTSGPs, into an applicator300may now be described with respect toFIGS.15A-E. Referring toFIG.15A, an applicator300, which may be used to collect and dispense the MTPs.

TPs is typically configured as a syringe which provides a well-known means to deliver and meter out controlled volumes.

The syringe applicator300also serves as a device to separate excess fluids from the MTPs. A cylindrical screen filter302is formed as an insert to the applicator body303and has an inner applicator chamber308lumen to receive a plunger304and piston306. Peripheral drain channels305may surround the filter302such that excess fluid within the MTPs may pass freely through the filter walls, through the drain channels305and out the drain outlets310. The screen filter may encompass 360° of the applicator or only a portion thereof, as shown inFIG.15A, so as to leave sufficient area to view the contents through a window307. The filter302may be a fine mesh, for example having 50 micron openings allowing fluid to pass through while containing MTPs. Alternatively, filter302, may be comprised of, for example, laser cut, woven mesh or acid etched perforated screens with specifically sized larger openings, so as to drain away smaller particles with solution, while selectively containing wetted particles larger than the utilized filter openings. The syringe applicator300could also be configured to separate liquid from the MTPs and dispense the MTPs onto or into a separate device or container for application, for example, into an attached syringe.

FIG.15Aalso shows that the processing device10is attached onto the processor44, for example, with a bayonet mount115. The processing device44may be cordless and include a low voltage DC motor114driven by a contained rechargeable battery. The battery is preferably recharged by connection to a remote ACDC charger. The low voltage DC motor may also be powered through the remotely connected ACDC power source. In both such manners the use of low voltage DC power enables the safe use of processing device10in the potential presence of an aqueous solution. The axially connected drive engagement46connects the motor shaft of the processor44to the central shaft assembly. Upon morcelization of the TPs, the motor114may automatically be slowed or stopped and raised so as to open the seal19below the impeller, causing the morcelized tissue mixture and solution to drain through the gravity driven drain39from the processing chamber through the outlet26and through the port320to enter the applicator300.

The applicator300is shown with the plunger304and piston306in its raised position and with the cap318in place to close the dispensing orifice. As the MTPs and solution enters the applicator300, the fluid is drained away from the MTPs as the fluid will freely pass through the filter walls302, through the flow channels305and exit the applicator300through the drain outlet310into the fluid waste drain container317. Thereafter, the plunger304may be advanced sufficiently into the applicator chamber308so as to enable the piston306to close off the port320. The applicator300may then be removed from the processing device10by disconnecting inlet port320of the applicator300from the outlet port26aof the processing device10. Thereafter, the cap318may be removed from the applicator300and a selected applicator tip301may be affixed to the dispensing orifice309for dispensing the MTPs in a manner which will be described in further detail hereinbelow.

A further embodiment of the present invention, shown inFIG.16, is similar toFIG.15Arelative to including a processing device10mechanically coupled onto a processor44. The drive engagement46may similarly be raised to open seal19to drain the container12through an outlet26, however, as shown inFIG.16(as well as in various earlierFIGS.1,2,3) the drive shaft42may include a rotor pump118, for example with fins integrally molded upon the drive shaft42. The rotor pump118is driven by the motor114through axis A, to circulate solution through an isolation device415.

FIG.16introduces a different isolation device415which circulates fluid from the processing device10through outlet26and inlet29conduits. Upon completing morcelization of the TPs, the motor114will be automatically raised, along with drive engagement46and drive shaft42, so as to open the chamber seal19below the impeller108, to release fluid and MTPs from container12.

The motor speed is changed, as appropriate, to pump the solution and TPs through the outlet channel26and through a diverter valve417to enter a cylindrical isolation chamber416containing a cylindrical filter tube402lining. The filter tube402may be, for example a woven mesh, perforated film or acid-etched screen with openings sized appropriately to capture particularly desired sized MTPs. The particles are captured within the isolation chamber416as fluid passes through the filter tube, and through circumferential drain channels405, exiting the isolation device415through inlet conduit29, to return into the container12of processing device10. Within several brief passes the motor114will automatically stop as the MTPs are substantially rinsed away from the container12and transferred into the isolation device415.

The diverter valve417may then be automatically or manually switched, for example turned 90° clockwise, to open a fluid path from the isolation chamber416into the applicator400. The inlet conduit29is positioned below the fill line (F) of container12, enabling the head pressure of contained fluid to substantially flush the MTPs from the isolation chamber416through the diverter valve417, through an applicator attachment411, and into the applicator chamber408of a detachable applicator400.

Thereafter, the diverter valve415is closed; the applicator400is disconnected from the applicator attachment411; and a plunger with piston (as shown for example in previousFIG.15B) is manually inserted into the lumen of applicator400. The cap418is removed (for example with a Luer type connection) and replaced with a selected applicator tip (as, for example introduced inFIG.15B). In this manner, the applicator400is ready to dispense the MTPs matrix to a desired autologous implant sight in a manner which will be described in further detail hereinbelow.

The processing device10, isolation device415and applicator400may be packaged as an integral sterile assembly. Applicators400may alternatively be sterile packaged separately.

Turning now toFIG.17, a further embodiment of the processing device10with processor44and applicator500is shown here coupled to a different type of isolation device515. In this embodiment a pump118will similarly circulate the solution with suspended MTPs from the container12, through outlet26and returning through inlet29both in fluid communication with an isolation chamber515which in this configuration employs cyclonic action to separate the MTPs from the solution. The system uses the principle of terminal settling velocity of solid particles in a centrifuge field. The outlet26, from the processing device10, enters tangentially into the isolation chamber516of the isolation device515. High velocity centrifuge fields within the hydro cyclone cause particles to migrate rapidly to the outside walls of the conical chamber516and will be forced to move downward on the inside of the conical walls through a valve517, through an applicator attachment511, and into the applicator500. A valve517may then be closed, the applicator500is disconnected from the collector, a plunger with piston is inserted into the applicator500, the applicator cap518is replaced with the dispensing tip of choice, whereupon the applicator is ready to dispense MTPs in a manner described hereinbelow.

A still further embodiment is shown inFIG.18where the processing device10is coupled to the processor44. InFIG.18, the inlet20and outlet channels26are shown in fluid connection with an isolation chamber616that employs a whirlpool like action to gather the swirling MTPs particles towards the central drain through which the concentrated MTPs will be deposited into an applicator chamber608within an applicator600. As with the above embodiments, the valve617is then closed, the applicator600is disconnected and the applicator cap618is replaced with a dispensing tip of choice. The applicator600is then ready to dispense the MTPs in a manner which will be described in further detail hereinbelow.

A still further embodiment is shown inFIG.19where the processing device10is coupled similarly as shown inFIG.16onto an isolation device716through an outlet26and an inlet29. Also similar to the embodiment ofFIG.16, the isolation device716contains an isolation chamber716, separated by a filter tube702from a drain channel or channels705, such that solution passing through the isolation chamber716will pass through the filter tube702, to pass through the drain channel705, to pass through the inlet29and be recirculated through the processing device10. However, in this embodiment, the outlet26and inlet29may include sealable closable ports720(not shown) such that the isolation device716may be detachable from the processing device10while containing fluid from leaking from the detachable outlet26and inlet29flow paths. In this manner the detached isolation device716may contain a plunger704and piston706and detachable cap718and together may be used as applicator700as similarly described inFIGS.15Cand D.

The schematically drawn circulating flow (CF) paths inFIGS.2and14have been significantly simplified, by not indicating the turbulent vortex swirl, so as to more clearly depict the recirculating nature of the fluid flow pattern. Early prototypes revealed that a vortex induced by the impeller, while desirable to continuously recirculate and mix the fluid suspended TPs, also caused the particles to travel many more circuitous times around the container12than necessary before being drawn through the morcelizing mechanism40and becoming morcelized.

FIG.20, an axial view, andFIG.21, a lateral view, introduce a preferred improvements to the processing device10to more expediently morcelize TPs. Placement of vertical baffle panels140, radiating from the stem28, effectively interrupt the single vortex. The baffles are positioned proximal to the apex of each set of converging blade edges122on the stationary cutting member102. The swirling fluid within the container12rebounds off each baffle140, creating a separate smaller vortex V adjacent to each baffle140. The smaller vortexes V carry the TPs (tissue particles) more expediently through each of the continuously closing breaches124of the morcelizing mechanism40.

Although the circulating flow (CF) paths within processing device10as described inFIGS.2and14represents a preferred embodiment (and is also included by way of example in multiple other FIGS.), it is not intended to limit the scope of the invention. Whereas an impeller108is employed to continuously recirculate fluid and TPs through the central volume (CV) of container12so as to repeatedly pass through the morcelizing mechanism, the fluid and TPs need not necessarily be recirculated through the peripheral volume PV upon return.FIG.22, therefore, teaches that the fluid and suspended TPs may be recirculated through the morcelizing mechanism40in other manners, by way of another example, to flow externally of the container12, through a recirculating conduit150.

Further, whereasFIG.22shows a rotor pump118, integral to shaft42and rotating about axis A, one skilled in the art would recognize that a fluid driving pump may alternatively be included elsewhere along a recirculating conduit150between an outlet26from the processing device10and a return inlet29to the processing device10. Further, as such, a circulating pump (not shown) need not be driven by or associated with a motor also used to drive the processor10and could be, for example, a separately operable fluid pump. Further, referring still toFIG.22, such a recirculating conduit150may include one or more diverter valves117, such that (upon completion of morcelization) the fluid and suspended MTPs can be diverted to circulate through any of various types of isolation devices, for example as described throughFIG.15A-E,16,17,18or19.

Further still, the impeller108ofFIG.22, used in a system configured with a recirculating conduit150, external to the container12of processing device10, need not have vanes configured for radial thrust110b. In such an embodiment, vanes with a pitch configured for axial thrust110aalone may be sufficient on an impeller108to facilitate circulation of the tissue bearing solution from container12along axis A, through outlet channel26, through a recirculating conduit150, and through inlet channel29to be continuously recirculated through the morcelizing mechanism40within container12.

A still further embodiment is schematically shown inFIG.23. Here a processing device910is shown used in conjunction with a centrifugal type of isolation device919to effectively separate and compact processed MTPs, as defined herein, for dispensing through a conventional syringe (not shown). The isolation device919may be shrouded within a protective cover976. The processing device910and isolation device919, together with enclosing shroud976, may preferably be integrated into a single unit, to be packaged and pre-sterilized as a single patient use as a disposable device as red bag medical waste. The device may be used multiple times within a procedure for an individual patient. The combined processing device910and isolation device919are configured to be axially aligned and fixably coupled, for example with a bayonet engagement960, onto an aseptically cleanable reusable processor944.

Upon completing the morcelization of TPs within a processing device910, the MTPs are released in solution through a drain939from the bottom920of container912. The drain939is preferably located about a central axis A of the processing device910or otherwise appropriately located on the bottom920of the container912, so as to fluidly communicate into a central chamber972of the centrifugal isolation device919.

One or more individual collection chambers973protrude radially from the central chamber972, each collection chamber having a distal outlet orifice974. The distal outlet orifice974has a standard threaded female Luer engagement for interchangeable attachment of a standard Luer cap975or a standard Luer tipped syringe (not shown).

The central chamber972and radially extending collection chambers973may be integrally formed as a hollow injection blow molded component, or produced as an assembly of injection molded components, or a combination, for example with injection molded Luer fittings affixed onto an injection blow molded unibody core.

The centrifugal isolation device919is configured to rotate at a high speed, for example up to 300 Gs, on precision radial type ball bearings (not shown), about an axis that is preferably coincident to or axially aligned with axis A of the processing device910. The spinning isolation device919is preferably encased within a protective shroud976. Should the Processing device910and the centrifugal isolation device919rotate about the same axis, the ball-bearing's inner shaft diameter may be sized sufficiently large as to enable independent rotation of the centrifugal isolation device919relative to rotation of the impeller908within the processing device910.

Upon being centrifugally spun for only a few minutes, the solution suspended MTP's will separate and become compacted within the radially extending collection chambers. The Luer caps975on the distally extending female Luer connectors974are then unthreaded and exchanged with appropriately sized standard syringes. Upon then drawing the compacted MTPs into the syringes, the filled syringes are disengaged from the isolation device919and a selected Luer fitting applicator tip (for example as previously described inFIG.15) is affixed, now ready for autologous MTP application to the intended site.

Morcelized Particulates

A solution of suspended MTPs, e.g., FTSGPs or other tissues particles as described herein, may be mixed in combination with other FDA approved additives, for example handling (i.e. in saline, a buffer solution, or BioLife Solution®, or other cell nurturing/preservation solutions, etc.). The mixture may be created within the processing device10, using the vortex circulation to achieve a heterogenous mixture of morcels comprised of naturally connected cellular and extracellular matrix material. Alternatively, some suspensions/dispersions of MTPs may be homogeneous. Whether the dispersion or emulsion produced is homogeneous or heterogeneous may depend on a number of factors, including without limitation, the type of tissue(s), the medium it is suspended in, the speed and temperature of the process, among other factors. An important advantage of the method of creating the MTPs suspension/dispersion of the invention, as well as the resultant suspension/disperisons per se, relates to the high cellular viability during and immediately after processing to achieve the morcelization. This ability to morcelized while maintaining such high cellular viability, as described herein, is unique to the present invention and not achieved by prior methods. The methods described herein produce suspensions or dispersions which contain MTPs having at least 50% viability immediately after processing, which is generally in real time at the bedside of the patient; or at least 60% viability immediately after processing; or at least 70% viability immediately after processing; or at least 80% viability immediately after processing; or at least 85% viability immediately after processing; or at least 90% viability immediately after processing; or at least 92% viability immediately after processing; or at least 94% viability immediately after processing; or at least 96% viability immediately after processing; or at least 97% viability immediately after processing; or at least 98% viability immediately after processing; or at least 99% viability immediately after processing. Generally, the processing may take about 1 hour, but desirably less than 1 hour, for example, 45 minutes or less, 40 minutes or less, 30 minutes or less, 20 minutes or less, or 10 minutes or less.

The MTPs may also be centrifuged to vary the density, viscosity and consistency of the tissue particles, as may be desirable for alternative surgical applications. Modulating the centrifuge speed and duration of centrifuging enables the customization of the resultant output tissue particle form, for example the consistency and density may present as a solution, or a paste, or a cream. Desirably, the resultant output (Id.) flowable and/or easily applied by spreading. The resultant output may further be presented, for example, as compacted tissue form or may be further spun to present as compacted cellular matter.

The MTPs as defined herein may be most efficiently delivered from a variety of fluid dispensing devices, most notably syringes, which are familiar and useful to easily meter controlled volumes. The targeted particulate sizes will pass freely as a fluid composition through the lumen of standard Luer connectors. Tips may be interchangeably attached onto an applicator, for example, with a standard Luer thread. A variety of interchangeable applicator tips may be included within a dispensing kit for selection as most appropriate for a specific application at the option of the surgeon for a given procedure. In a cream, paste or fluid form, the MTPs, for example FTSGPs, may be dispensed from a syringe through various selected tip types of applicator tips. A tip may have a narrow/long fanned outlet orifice to spread over a large area. Such a fanned tip may be comprised of a flexible low durometer silicone or thermoplastic elastomer and may have a thin flexible edge, so as to be useful to gently and evenly spread the MTPs over large and/or irregular wound surfaces, for example burns.

In a dense form, the MTPs of the invention may be spread or applied over areas or into crevices, for example with a spatula. In a dense form, the MTPs, for example Cartilage Particles (CPs), may be used as a filler, for example for cartilage defects. As such the MTPs, such as CPs or other morcelized tissues particles as defined herein, may be mixed with fibrin glue, autologous Platelet Rich Plasma, growth factors or other FDA approvable materials, for example as a binder. Cartilage or organ MTP's may also be delivered through an endoscopic syringe attachment.

In a fluid a cream or solution form partial thickness dermal skin graft particles may alternatively be dispensed from a syringe, through a flexible cannula or a needle, for subdermal applications, for example, to fill cosmetic defects. For delivering the various MTPs as described the lumen may range, for example, from 22 to 18 gauge, or most notably 22 or 21 gauge.

In a highly soluble or dispersible form the MTPs, which in such a case may be of the smaller variety, as described herein, may be sprayed over large areas. In the case of MTPs for use on burns or open wounds, a non-adherent surgical wound dressing, may be used to prevent the applied MTPs from migrating while keeping the wound site moist and protected from infection. Such commercially available dressings include, for example, commercially available Drawtex, Sofsorb, Kalginate, or Aquasorb dressings.

System Methodology

A system of devices is described to perform a process in a methodical sequence. With reference additionally toFIG.24, which shows the system progression in operation, the system and process includes: 1) a processing device950is used to introduce and morcelize tissue into tissue particles (MTPs) within a solution; 2) an isolation device is then used to separate for dispensing morcelized tissue particles (MTPs) suspended in solution from the preponderance of solution; and 3) an applicator device952is then used to surgically dispense and apply the collected morcelized tissue particle matrix. Heretofore we have described several alternatively configured devices and methods employing non-limiting details with which to accomplish a three-step process to achieve the desired outcomes.

A processor device may be used in conjunction with various types of isolation devices. For example, an isolation device may take the form of a device including a filter tube through which solution flows to isolate MTPs as described inFIG.15A-E,16and19; or an isolation device utilizing cyclonic action within a chamber to isolate MTPs as described inFIG.17; or a device using a whirlpool action within a chamber as described inFIG.18; or a device performing as a centrifuge as described inFIG.23; or the MTPs may simply be isolated by sifting the preponderance of solution away through a screen (not shown); or settled MTPs may be drawn from the solution using a standard syringe; or any number of other methods may be contemplated to isolate MTPs in suspended from the preponderance of the solution.

A processor device may also be used in conjunction with various types of applicator devices. For example, an applicator device may be a standard syringe; or an isolation device may additionally be deployed for use as an applicator as described inFIG.15A-Eor19: or a spatula may be used to manually apply the MTPs matrix; or the tissue particulates in liquid suspension may be sprayed over large wound areas; or any number of other methods may be employed to deliver and apply MTPs in a controlled manner.

FTSG Process Verification Studies

Full Thickness Skin Grafts (FTSGs) harvested from a human abdominoplasty were prepared in accordance with the methods disclosed herein and using the apparatus and systems disclosed herein.

Harvested sample FTSGs of various noted sizes were each separately and individually placed into fabricated experimental test apparatuses modeled as generally described inFIGS.1-4without baffles andFIGS.20-21, with baffles. The processing devices were filled with 35 ml of buffered saline solution, pre-chilled with ice chips. The FTSGs were then morcelized by subjecting the samples to a slicing speed of approximately 550 rpm for incrementally stepped durations, timed in minutes at ambient room temperature of 70° F. Earlier tests demonstrated insignificant temperature rise of the chilled buffered saline water over the lapse time processing each sample.

MTPs were quantitatively assessed to determine cell viability. Processed FTSGPs suspended in solution were transferred using a syringe into 0.5 ml aliquots. The aliquots were maintained chilled in a container of chipped ice. The samples were spun down in a centrifuge at 700 RPM for five minutes followed by removing the supernatant. Quantitative cell viability analysis was performed using standard trypan blue test protocols to stain and count living cells versus purple ruptured dead cells. A table included asFIG.25documents highly viable cellular viability test results ranged consistently between 87% to 98% viability—across multiple exemplary sample lots and processing parameters and processing durations, a sampling of which are described below.

Additional MTT tests yielded similar quantitative results to consistently confirm data reliability. MTPs were also qualitatively assessed. Resultant morcelized tissue particles, suspended in solution, were drawn from the processor through a cannula into a syringe and expelled into a petri dish to form a shallow pool or puddle aside a metric scale. Photos of the morcelized FTSGPs are shown, herein, each photo identified by test sample numbers, to visually and qualitatively document relative morcel sizes and particle appearance. As evident in the images, tissue particle sizes are relatively consistent within each sample. Maximum particle sizes became progressively smaller with longer total duration of processing time.

In exemplary morcelization studies, ten portions of full tissue skin grafts (FTSGs), each approximately 12 mm×6 mm×4 mm thick were morcelized in a processor without baffles, containing 35 ml of buffer solution with the blades rotating at approximately 550 RPM.FIG.26A(test 1b) shows a sample of morcelized FTSGPs in fluid suspension drawn from the processor after 4 minutes of processing. FTSGPs in solution were transferred to form a shallow pool in a petri dish to visualize individual particles. The maximum sizes of individual particles appear to generally be no more than approximately 1.5 to 2.0 mm on any axis. The majority of particle sizes appear less than 1 mmFIG.26shows a subsequent sample then drawn after an addition three minutes, for a total of 7 minutes. The second sample appears more densely populated with particles and the typical maximum particle sizes appears to have been reduced to no more than 1.5 mm across.

In a next exemplary morcelization study, still using the same patient tissue, the processing chamber included three baffles and four (versus 10) portions of FTSGs. The sample was again processed with 35 ml buffer and 550 RPM. As shown inFIG.27, after being processed for 3 minutes, the resultant morcelized FTSGPs were similarly sized and particle density as the previous sample after 4 minutes.

In other studies, not included here, similarly morcelized FTSGPs have been demonstrated to be injectable through 22 gauge needles. The term injectable as used herein is meant to include dispensing through a syringe and is not intended to be limited to being injected only into the body, but also includes dispensing onto the body, such as onto a wound.

In a next exemplary morcelization study (test 4a)—again using the same patient tissue and processing device and process parameters—multiple larger portions of tissue, measuring approximately 2 cm×3 cm were inserted into the processor and morcelized for 4 minutes.FIG.28Ais a close-up of a portion of the FTSG prior to processing, with a sectional view revealing the thin layer of epidermal tissue (typically including pigmented stratum corneum, stratum lucidum, statum granulosum, thickly cell populated stratum spinosum, and stratum basale), over the thicker layer of generally white dermis (including dermal papilla, stem cell rich hair follicles, sweat glands, capillaries, sensory nerve fibers, sebaceous glands and other dermal components—all contained within an abundance of collagen fibers and connective tissue).

FIG.28B(still test 4a) shows the resultant dense mixture of morcelized particles (MTPs) suspended in 35 ml of buffer solution, contained within the processor chamber.FIG.28Cshows an enlarged view of the densely populated tissue particle solution presented in a petri dish.FIG.28Dshows an enlarged view of the processed FTSGPs, annotated to point out that the mixture contains differing amounts of epidermis (pigmented) versus dermis (generally whiter), varying proportionally as anticipated looking at the sectional view of pre-processed tissue. It also appears that the epidermal tissue (more densely populated with cellular structure) slices more sharply, relative to the more fibrous dermal tissue. The epidermal and dermal tissue particles appear to be distributed rather consistently throughout the mixture.

FIG.29Awere processed using the same device and same process parameters asFIGS.28A-D, however, on a different day and with abdominoplasty derived tissue from another patient. Together, these studies demonstrate a repeatable process, able to achieve consistent FTSGPs outputs, relative to qualitative appearance and particle size, as well as, consistently high quantitative cellular viability.

A similarly sized (slightly larger) single portion of FTSG, measuring approximately 2.5 cm×4.5 cm was morcelized for 4 minutes before taking the photo forFIG.29A(a different sample test 1b).FIG.29B(sample test 1c) was then morcelized for an additional 3 minutes, for a total of 7 minutes. AndFIG.29C(sample 1d) was morcelized an additional 3 minutes, for a total of 10 minutes. Only a small shallow puddle of resultant FTSGPs is shown in each of these images so as to better visualize individual particle sizes. The overall volume of processed FTSGPs for this study appeared as densely populated as in the previous study forFIG.28A-D.

The tabled data inFIG.25demonstrates relative consist and repeatable cellular viability outputs for each of the FTSGPs mixtures documented for exemplary test samples included inFIGS.26A-B,27and29A-E.

FTSGPs shown inFIG.29C(test 1d) above and a subsequent sample (test 2b) were further centrifuged in 1.5 ml aliquots for 4 minutes at 700 RPM. The resultant tissue form, shown inFIG.29Ddemonstrates the ability to achieve a fine paste-like mixture which can be dispensed through a syringe as demonstrated inFIG.29E. Such a FTSGP tissue form may be easily applied and dispersed, for example, over expansive wound surfaces.

Articular Cartilage Process Verification Studies

Articular cartilage was harvested from the peripheral edges of a bovine knee condyle using a 2.5 mm ring curette and then morcelized in accordance with the methods disclosed herein and using the apparatus and systems disclosed herein.

The harvested cartilage portions, shown inFIG.30A, (test 4a on Jan. 16, 2017) ranged in approximate size from about 1.0-2.2 cm long, 2-2.5 mm wide and 0.75-1.2 mm thick. The portions of cartilage were inserted 3-4 at a time into 35 ml of buffered saline solution, within an apparatus as described previously inFIGS.1-4without baffles, with the slicing blades rotating at about 550 RPM within the morcelizing mechanism. The cartilage tissue grafts were morcelized (MTPs) for a total duration of 15 minutes at room temperature.

The resultant cartilage morcels are shown inFIG.30B, demonstrating the ability to also finely morcelize articular cartilage within a device and by a process as described herein to similarly process full thickness skin graft tissue.

Further details of the present invention are shown and described hereinbelow with respect toFIGS.31-38. These details include the technology advantages and components, the needs and benefits, the technology procedure, tissue types and preparation, the process, variable tissue particle sizes, tissue dispensing options, clinical indication and development status.

It is contemplated that the present invention meets a significant unmet need. Full thickness skin grafts are the gold standard for chronic wounds and burns, but are rarely used because dermatomas create donor sites that do not heal and the procedure must typically be done in an operating room.

The present invention can generate a full thickness skin graft rapidly without leaving a conventional donor site to heal.

The present invention can also be customized to be applied to fit wound anatomy and can be done as an office procedure. The resulting process of the present invention and the grafts produced thereby are fast to process, are minimally invasive, antiseptic, provide superior viability and are cost effective solutions for wound healing. The system, equipment and process of the present invention can be conducted at bedside, including preparation of the morcelized TPs (MTPs) and formation of a fluid having a pH to help sustain the tissues, and dispensing of the MTPs onto/into the area intended to be treated, which may be a wound, a cosmetic or plastic surgery area, an internal organ area and the like.

As shown inFIG.31, use of the device of the present invention, which includes a processing device950, an applicator952and reusable equipment954, allows for retention of the original tissue structure, high tissue/cell viability (90-95%) and the ability to vary tissue particle size. In addition, versatile dispensing methods such as spread paste, spray and injectables may be used. These are all acceptable for in-office procedures and may be completed within approximately 20 minutes or less, desirably about ten minutes or less. Moreover, the present invention allows for processing of multiple tissue types such a skin, cartilage and organs.

Referring toFIG.32, the complete total procedure is completed within thirty minutes. Preparation955is improved as the procedure results in fast healing, low pain levels, fast harvesting and processing, and a suture closed donor site. The processing956to form the MTPs is conducted in a closed antiseptic system taking no more than about ten minutes. The process is automated and can accommodate variable particle size and results in high cell viability (90%+, such as 99%). Application957may be done by selectable tips on irregular surfaces and with variable wound sizes. Also, the application may be injectable. The MTPs of the present invention are desirably prepared in a pH suitable for maintaining viability once they have be morcelized into the intended sizes. The fluid containing the morcelized highly viable MTPs may be dispensed using a conventional syringe onto or into the area to be treated. The fluid containing the MTPs suspended therein may be applied to a wound, or other area of the body in need of treatment, such as in a body joint, a plastic surgery application or cosmetic application, or other area of use to enhance the health the of tissue and/or overall appearance and health of the patient.

Turning now toFIG.33, therein as shown, the preparation process using both full thickness skin grafts (FTSGs) and cartilage grafts (CG).

FIG.34shows the basic three-step process with respect to morcelized abdominoplasty tissue in solution dispensed from a 1 mm syringe. This includes introduction958, morcelization (cutting the donor site tissue in particles)959, and dispensing961.

FIG.35shows variable tissue particle sizes, which may be formed by the inventive process and using the devices and systems discussed herein, of full thickness skin graft particles (FTSGs) (also referred to as morcels or MTPs) containing cells and extracellular components.

FIG.36shows various dispensing tip options and devices including spreading980using a fan-tip wiper981; a paste982using a cannula983; spray984using a nozzle985and an injectable986using a needle987, all coming from an appropriate applicator device952.

FIG.37shows non-limiting examples of clinical indications including wound healing990, skin anesthetic injectable991and cartilage repair992using injection devices with appropriate tip selection.

FIG.38shows morcelized tissue particles (MTPs) of bovine knee articular cartilage, as well as morcelized tissue from abdominoplastic formed using the inventive process. These morcelized cartilage and skin particulates (particles) may be disbursed to a patient using any of the dispensing devices described herein. These results had been repeatedly verified to have 87-98% cellular viability using standard tripan and MTT test protocol.

Further embodiments of the present invention are now described with respect toFIGS.39-50where similar description and reference numerals are used to describe similar components.

FIG.39shows a processing device1010having a processing container or chamber1012which is similar to that described above. In addition, processing device1010also includes a filter chamber1100, a drain chamber1200and a dispenser1300. The processing chamber1012is filled with a measured volume of sterile solution, preferably sterile saline. The TBS is inserted into the processing chamber1012through an upper open end1014and into the solution. The sterile solution and TBS is shown collectively as tissue fluid (suspension)1018. The processing chamber1012may include a cover or lid1016for closure of the open end1014. The lid1016may be detachable or preferably hinged upon the processing chamber1012to cover the open end1014during processing.

In the present embodiment, the morcelizing mechanism1040is driven by an axially rotating drive shaft1042that passes through a suitable rotary shaft seal1047that may be a silicone part held in place by a retaining clip1048that prevents fluid from leaking from the processing chamber1012.

Processing chamber1012further includes baffle panels1050within the interior thereof. With additional reference toFIGS.49and50, the baffle panels1050, arrayed about a center axis, are preferably integral with the interior wall1013of the processing chamber1012or may adjacently abut the interior wall. The baffle panels1050enhance flow and circulation characteristics of the MTPs (e.g. FTSGs) within the fluid through the morcellation mechanism1040, by disrupting otherwise circumferential flow to effectively divert flow into and through the morcelizing mechanism1040.

Referring again toFIG.39, fluid communication is provided between processing chamber1012and filter chamber1100through a connecting channel1416. A flow valve1417positioned within connecting channel1416is shown in the closed position. The valve could be a ball valve, choke valve, stopcock or any other means to controllably open/close fluid flow from the processing chamber1010into the filter chamber1100and further on into the drain chamber1200.

The drain chamber1200mentioned above is shown as an annular shaped container.

FIG.40shows the processing device ofFIG.39with the flow valve1417in the open position. Fluid communication between the processing chamber1012and the filter chamber1100through connecting channel1416enables the fully processed MTPs in fluid suspension1019to drain into the filter chamber1100. Upon completing morcelization, as discussed above, the morcelizing mechanism1040continues to rotate to maintain the MTPs in suspension as the MTPs and fluid1019drain through the opened valve1417and into an inner chamber1410of the filter chamber1100indicated by arrow A.

The drain chamber1200can include a vent (not shown) to enable air within the head space to escape ahead of incoming fluid flow when the flow valve1417is opened.

Additionally and as more fully shown inFIG.48, within the filter chamber1100, a cylindrical filter1402separates the inner chamber1410from an outer chamber1420. Ribs1430protruding from the inner surfaces of the walls of filter chamber create linear drain channels1435and also support the cylindrical filter1402. The structure of the cylindrical filter may be interrupted locally for flow from the connecting channel1416into the inner chamber1410of the filter chamber1100. Alternatively, the connecting channel1416may pass over the filter1402into the inner chamber of the filter chamber. The outer surface of the filter1402is supported by the array of vertical ribs1430that run lengthwise between the cylindrical filter1402and inside the outer filter chamber1420. Spaces between the ribs1430form the drain channels1435that enable the filtrate1023to pass through the filter and drain down the peripheral drain channel1435into the drain chamber1200below the filter chamber1100as indicated by arrows B inFIG.40. The drain channel1435may alternatively drain the filtrate to a dispensing port1415to void or collect the filtrate externally from the device.

The cylindrical filter1402, which may or may not form a complete 360 degree cylinder, permits fluid and particles smaller that the filter pore size (mesh size) to drain through the filter and down the drain channels1435and out through a lower filter chamber drain port1440(FIG.40) communicating with the drain channels1435into the drain chamber1200.

The filter1402mentioned above can be formed into a cylinder from a flat filter perforated sheet such as a stainless-steel sheet with photochemically etched openings or for example lasercut perforations through a plastic sheet. The cylindrical surface area of the filter is large enough and with sufficient pore density to prevent MTPs from fully occluding the filter. MTPs settle and collect on the inner surface of the filter and settle on the inner concave bottom1411of the filter chamber1100. The filter1402, separates the inner filter chamber1410from the outer filter chamber. The filter1402serves as a sieve to collect MTPs which are larger than the filter perforations and to expel excess solution and filtrate particles smaller than the perforations from the mixture.

A piston1306passes through the lumen of the inner filter chamber1410and is attached to a plunger1304which extends upward and through a filter cap1308. The cap1308and the filter chamber1410are mechanically secured via threads, although could be secured by various other means such as snap features, plastic welds, or adhesive.

The drain chamber1200may or may not form a 360-degree annular enclosure. The drain chamber1200may be any type of container or means of collecting the filtrate. An absorbent, such as a compound or an absorbent material, may be included inside the drain chamber to congeal, coagulate or otherwise increase the viscosity of the filtrate.

Referring more specifically toFIG.41, the processing device1010ofFIGS.39and40is shown but with the plunger1304in the down position and the filtered MTPs1021transferred into the dispenser1300. The downward movement of the plunger1304(indicated by arrow C) causes the piston1306to transfer MTPs1021that have adhered to the inner wall of the filter1402and onto the inner concave bottom1411of the filter chamber1100where other MTPs have also settled. As the piston1306advances through the filter chamber1100and approaches an inner concave bottom1411. MTPs are transferred from the filter chamber1410, through a dispensing port1415and into the receiving dispenser1300. As MTPs are driven by the piston1306into the receiving dispenser1300, a syringe plunger1360within the dispenser1300will be displaced outward from the dispenser. The port1415may be a standard type threaded Luer connector. The dispenser1300may be a standard type syringe1301attached onto the dispenser port1415of the filter chamber1410or any other means to collect the MTP matrix. The dispenser containing the MTPs matrix is removable for dispensing the MTPs, for example onto the treatment site.

Referring now toFIG.42, an embodiment similar toFIG.39is shown where the flow valve is replaced instead by a simple connecting channel1450between the processing chamber1012and the filter chamber1100. The head pressure of fluid1018within the processing chamber1012is initially unable to drive the fluid through the connecting channel1450, constrained in a state of equilibrium upon the air entrapped below within the interconnected filter chamber1100and the drain chamber1200.

Similarly to the embodiment ofFIGS.39-41, the embodiment ofFIG.42, also includes a piston1306, attached to a plunger1304that passes through a filter cap1308. However, here, a plunger seal1307within the filter cap1308prevents air within the filter chamber1100and drain chamber1200from escaping to atmosphere alongside the plunger shaft1304. The seal1307may be an elastomeric disk, for example low durometer silicone, with a hole through which the plunger passes and is captured and held in compression, between the filter cap1308and chamber1100. The cap1308is secured onto the filter chamber1410, for example via threads, snap features, plastic welds, or adhesive.

A vent valve1452is positioned in the upper surface of the drain chamber1200. The filter chamber1100and the drain chamber1200maintain an airtight enclosure while the vent valve1452is spring-loaded closed. Fluid is contained by the processing chamber1012with a fluid level that is above the connecting channel1450that communicates between the processing chamber1012and the filter chamber1100. The sealed filter chamber1100, drain chamber1200and connecting channel1450maintain a static volume of air that prevents the fluid1018within the processing chamber1012from flowing into the filter chamber1100. The connecting channel1450restricts fluid communication between the processing and filter chambers such that fluid and air cannot flow through the connecting channel1450while the device is at rest and the vent valve1452is closed in its static position.

FIG.43shows the same device as inFIG.42only the vent valve1452on the drain chamber1200is now shown open, enabling head pressure of the fluid within the processing chamber1012to drive air inside the filter and drain chambers1100and1200to exit out through the vent valve1452. In this manner the expelled air is displaced by the fluid entering the filter chamber, as the MTP are collected within the filter chamber, and the sieve function of the filter enables the fluid and filtered filtrate1023to then flow further to be collected in the drain chamber.

FIG.44is similar to theFIG.42embodiment showing that the air pressure inside the drain chamber1200and the filter chamber1100similarly prevents the tissue fluid1018in the processing chamber1012from draining into the filter chamber1100until a vent valve1454is opened. The vent valve1454inFIG.44is shown in a closed state. This vent valve1454is configured to be actuated from the bottom of the device such that the opening of the valve1454can be automated electromechanically. A valve seal1456is located inside the upper region of the drain chamber, well above the maximum level that enters the drain chamber.

The device ofFIG.44is shown inFIG.45with the vent valve1454in the open position. An actuator shaft1458is shown extending upward from an operatable processor1480(shown in dotted lines), to drive the vent valve1454into the up/open position. The MTPs1019have drained into the filter chamber1100and the filtrate1023has passed through the filter1402and drained into the drain chamber1200.

FIG.46shows a processing device similar toFIG.44in that it is configured to be actuated from below from the operatable processor1480. This figure shows the device configured to provide for automation of a linearly actuated flow valve1490between the processing chamber1012and filter chamber1100, as well as a linearly actuated plunger1492within the filter chamber.

In this embodiment, the linear actuated valve1490would be closed (FIG.47) in the initial state when fluid and TBSs are contained within the processing chamber for processing. As shown inFIG.46, the valve1490is then driven to an open state by an actuator shaft1491following morcelization, to enable fluid to drain into the filter chamber1100and pass through into the drain chamber1200, while the MTPs1019drain into the inner filter chamber1410and settle above the piston1493.

In this embodiment, the linearly actuated piston1493is positioned in the bottom of the filter chamber1100during processing, as well as upon opening the flow valve1490. With use of the automated operable processor1480, upon fully emptying the processing chamber, the motor drive in the operatable processor1480will be automatically stopped and the piston1493will then be automatically raised to drive the MTPs captured within the filter chamber1100up into a detachably affixed dispenser1300.

A linearly positionable piston raising rod1492may be adjoined by a seal (not shown) to prevent leakage from the filter chamber1100. The flow valve1490and/or the valve actuating rod may similarly be adjoined by a sealing means to prevent leakage from the connecting channel1450.

FIG.47depicts a detail of the processing device inFIG.46with the piston1493fully translated to its second state transferring the MTPs from the inner filter chamber1410into a dispenser1300. The dispenser1300can be removed to dispense the MTPs1021.

Referring again toFIGS.49and50, the baffle panels1050mentioned above are further shown. The morcelizing mechanism1040is visible in the cross section ofFIG.49. The baffle panels1050enhance the flow patterns within the processing chamber and increase the recirculation of MTPs through the morcelizing mechanism. Preferably there are three baffle panels, however, there could be more or less. The baffle panels1050ofFIG.49are shown preferably integral to the processing chamber wall1013, facilitating blended surface transitions for effective fluid flow and to avoid such sharp edge features as might otherwise cause circulating MTPs to become hung-up. Alternatively, however, the baffle panels1050could be separate from the wall1013and may have space between the processing chamber wall1013and the baffle walls1050(as shown, for example, in earlierFIG.20). The baffles of the panels1050can vary in shape, height, and geometry, and may have perforations or openings in the baffle panels1050.

Further embodiments of the blade/cutting assembly1699used in the morcelizing mechanism1040of the present invention are shown inFIGS.51-57.

FIGS.51,51A and52show a rigid blade1700that is held in compression by a spring1710(Belleville washer) to a cutting disk1720and a pin1730using a retaining clip1740. The blade1700has sharpened cutting edges1711on opposite ends of the blade such that both cutting edges are in the direction of rotation. The cutting edges1711preferably are straight but could alternatively be curved within the SP. An impeller1717drives the rotation of the blades. Vertical contact surfaces1719on the impeller1717engage with rear vertical flat engagement surfaces1721on either side of the blade1700to drive the blade rotationally about a central axis as the impeller1717rotates.

The blade1700has upper flat surfaces1712that ride in the SP and against the disk. The upper flat surfaces1712could also be offset at an angle relative to the SP, creating a relief angle, so that only the edge of the blade is in the SP and in contact with the disk1720.

The cutting disk1720has preferably three cutting apertures or breaches1760which are radially arrayed about a central axis. The blade1700preferably includes two opposed cutting edges1711projecting radially outward about a central axis. The blade1700is mounted about the axis pin1730, such that the cutting edges1711of the blade1700are in the shear plane (SP) side of the cutting disk1720.

The cutting disk1720is a stationary cutting member similar to that described above. A mating flat or flats1731on the axis pin1730and on the central axis hole1722through the disk1720prevent the disk from rotating upon the axis pin. An enlarged diameter step1732on the axis pin1730controls axial movement of the disk upon the axis pin. Similarly, the pin has two opposing flat surfaces1734that mate into keyed surfaces in a central hole within the processing chamber. The retaining clip1740, for example an ‘E-clip’ or ‘C-clip’, is engaged onto an annular groove about the axis pin and controls the axial movement of the blade1700upon the axis pin1730while leaving the blade free to rotate upon the pin1730.

One or more washers1733may be placed between the blade1700and retaining clip1740to prevent the retaining clip1740from being dislodged as the blade rotates, as well as to prevent the retaining clip from wearing against the rotating blade1700. One or more springs, for example Bellville washer1710or a wave washer, are positioned between the blade1700and washers1733, axially opposed between the retaining clip1740and the disk1720, against the step on the axis pin1732. The blades1700are secured in compression against the shear plane (SP) side of the disk1720.

Another embodiment of the blade/cutting assembly1699shown without the impeller inFIGS.53-54is a combination of the morcelizing mechanism40inFIGS.5-12and the morcelizing mechanism1040inFIGS.51-52. The rigid beam1800ofFIG.53-54supports two blades1810(or cutting blade members) on either side of the rigid beam and replaces the blade1700inFIG.51. Each blade1810includes a cutting edge1811. As shown in the blade assembly1699inFIGS.53and54, the rigid beam1800and supported blades1810are held in compression against a cutting disk1720by a spring1710held together by a retaining clip1740and a pin1730that keys into the disk1720. The blade retaining features of the rigid beam1800inFIGS.53-54are similar to the blade retaining features on the impeller108inFIGS.5-11. The rear portion1815of the blade1810, opposite of the cutting edge1811, is secured to the rigid beam1800by screws1817in lieu of a slotted feature in the impeller108shown inFIGS.5-7andFIGS.9-11. The blade could be secured to the rigid beam by other means such as a slotted groove, heat stake, a pin or pins. The rigid beam1800and blades1810are driven rotationally against radially facing flats1821by an impeller as described above. The blades1810may also be mounted to the rigid beam1810such that they are parallel and flush against the SP surface of the disk1720. The blades1800preferably have a straight edge although the edge could be curved, or circular. The blade edges1811are held against the SP of the disk1720in compression.

FIG.55depicts a blade assembly similar to that shown inFIGS.53-54, except blades1811are thin and can elastically deform or bend in compression against the cutting disk1720. The blade, being slightly flexible, may bend into the aperture openings1760in the cutting disk1720such that the cutting edges of the blade ride against the cutting edges of the cutting disk.

FIG.56shows output of a FTSG processed in accordance with the present invention for 3 minutes into approximately 1 ml output volume of morselized FTSGPs, with each morsel generally 1-3 mm in size. Smaller micro-particle filtrate has been filtered out of the mixture. For scale reference, the morselized FTSGPs ofFIG.56are shown within a 9 cm petri dish alongside a metric scale. The morsels are shown diluted in solution to distribute the morsels over the area within the petri dish.

FIG.57shows half of the total 1 ml output volume of FTSGPs ofFIG.56, diluted in solution within a 9 cm diameter petri dish. The morsels are again generally spaced apart over the area of the petri dish, in this example illustrating a distribution of morsels at 50% density relative to the full output volume shown inFIG.56.

FIG.58shows a quarter of the total output volume of FTSGPs ofFIG.56, again diluted in solution within a 9 cm diameter petri dish. In this example, the generally spaced apart morsels illustrate a distribution of morsels at 25% density relative to the full output volume shown inFIG.56.

Referring now toFIG.59, an excised elliptically shaped FTSG, is shown here for scale reference in a 9 cm diameter petri dish (shown in partial view). Placed upon a 1 cm scale grid, the excised FTSG is seen to be approximately 1 cm×2 cm.FIG.60shows an abundance of FTSGPs, also in a 9 cm diameter petri dish, processed for 4 minutes from the FTSG ofFIG.59in accordance with the present invention.

FIG.61shows a larger excised elliptical FTSG measuring approximately 5 cm×2.5 cm, relative to the scale below.FIG.62shows an exponentially larger output volume of FTSGP, similarly processed for 4 minutes, from this larger FTSG. The compared FTSG input and FTSGP output volumes shown inFIGS.59and60, relative toFIGS.61and62, illustrate, by way of example, how a desired output volume of processed FTSGPs is easily achieved by varying the input volume of FTSG.

FIG.63shows finely morselated cartilage. Varying the processing time, for example from 4 minutes to 12 minutes, enables precise and reproducible control of output morsel size. In this example, a small biopsy sample of cartilage has been processed to output an abundance of small morsels, allowing for broad area coverage. Precisely reproducible small morsels may be delivered by syringe or through an arthroscopic portal.

FIG.64shows a planar slice through a single particle of morselized cartilage using fluorescent confocal microscopy. Staining with calcein AM reveals viable cells as green and dead cells as red. Both green and red are shown together here in this grayscale image. Cartilage morsels contain highly viable tissue components. The live/dead ratio of this exemplary scan is 88%.

The above-presented description and figures are intended by way of example only, and are not intended to limit the present invention in any way except as set forth in the following claims. It is particularly noted that persons skilled in the art can readily combine various technical aspects of the elements of the various exemplary embodiments described above in numerous other ways, all of which are considered to be within the scope of the invention.