Patent Description:
The transfer of adipose tissue to various regions of the body is a relatively common cosmetic, therapeutic and structural procedure involving the harvest of adipose tissue from one location and re-implantation of the harvested and, oftentimes processed tissue, in another location (see Coleman <NUM>; and Coleman <NUM>). While being largely used for repair of small cosmetic defects such as facial folds, wrinkles, pock marks and divots; the transfer of adipose tissue has recently been used for cosmetic and/or therapeutic breast augmentation and reconstruction (Bircoll and Novack <NUM>; and Dixon <NUM>), and augmentation of the buttocks (Cardenas-Camarena, Lacouture et al. <NUM>; de Pedroza <NUM>; and Peren, Gomez et al.

In the past, adipose tissue grafts and methods of adipose tissue transfer have been plagued with difficulties and side effects including necrosis, absorption of the implant by the body, infection (Castello, Barros et al. <NUM>; Valdatta, Thione et al. <NUM>), calcifications and scarring (Huch, Kunzi et al. <NUM>), inconsistent engraftment, (Eremia and Newman <NUM>), lack of durability, and other problems arising from lack of neovascularization and necrosis of the transplanted tissue. One of the biggest challenges in adipose tissue transfer is absorption of the implant by the body and volume retention of adipose tissue grafts following transfer. When adipose tissue is harvested or washed, the space between individual pieces of harvested adipose tissue is filled by liquid (e.g., water, blood, tumescent solution, oil). When this tissue/fluid mixture is implanted into a recipient the liquid portion is rapidly absorbed by the body resulting in loss of volume. The process by which the amount of fluid is removed from the tissue/fluid mixture is frequently referred to as "drying the adipose tissue" or "dehydrating the adipose tissue". The content of red and white blood cells and the like within an adipose tissue graft can also significantly affect the volume of graft retained after graft transplantation, due to induction or exacerbation of an inflammatory response. Another aspect of tissue retention relates to the amount of lipid within the adipose tissue graft. It understood that the presence of free lipid (meaning lipids released from dead or damaged adipocytes; also referred to as oil) in adipose tissue grafts can result in induction or exacerbation of an inflammatory response with substantial phagocytic activity and consequent loss of graft volume.

It is also known that mixing unprocessed adipose tissue with a concentrated population of adipose-derived regenerative cells overcomes many of the problems associated with adipose tissue grafts and adipose tissue transfer, as described above. Specifically, supplementing unprocessed adipose tissue with concentrated populations of adipose-derived cells comprising adipose-derived stem cells increases the weight, vascularization, and retention of fat grafts. (See <CIT> and co-pending <CIT>). Adipose tissue fragments supplemented, or mixed, with a concentrated population of cells including adipose-derived stem cells exhibit improved neoangiogeneis and perfusion in grafts when compared to unsupplemented grafts of adipose tissue alone in animal models. Further, adipose tissue grafts supplemented with adipose-derived regenerative cells that comprise adipose derived stem cells show increased graft retention and weight over time, when compared to unsupplemented grafts. (See <CIT>). Further, the processing of adipose tissue in a closed, sterile fluid pathway greatly reduces the chance of infection. The improvement in autologous transfer of adipose tissue seen in the animal models described above has also been replicated in human clinical studies. Nevertheless, the isolation and purification of concentrated populations of adipose-derived regenerative cells comprising adipose-derived stem cells (ADSCs), usually involves a series of washing, digestion, filtration and/or centrifugation steps, which can reduce the yield of viable cells, require mechanical equipment and specialized clinicians, and/or can compromise the quality, appearance, longevity, hydration or efficacy of the graft.

Additionally, stresses could cause undesirable reactions to harvested adipose tissues. Such stresses include, for example, exposure to environmental pathogens, which are mentioned above, and prolonged post-harvest storage, etc. Therefore, there is a need for in-situ harvest, cleaning, and use of an adipose tissue graft for biomedical applications.

The need for additional approaches to prepare and optimize adipose tissue grafts and implants and to isolate and/or concentrate adipose-derived regenerative cells is manifest.

<CIT> describes methods of manipulating subdermal fat in a treatment area. Such methods include inserting a needle (e.g., a micro-coring needle) through the dermis to the subdermal fat layer in order to excise a portion of tissue from the treatment area. These insertions result in a portion of the dermis and a portion of subdermal fat to enter into the needle. Removal of the needle results in excision of the portion of dermis and subdermal fat that entered into the needle.

<CIT> describes spatulated cannulae for use with negative pressure or liposuction equipment provide a myriad of benefits including reduced recovery time, improved procedural efficiency and others. A spatulated cannula generally comprises a planar spatula tip disposed at a distal end of a cannular body.

<CIT> describes that adipose tissue containing microvascular endothelial cells can be harvested using a collection apparatus incorporating an elongate cannula having apertures with tissue cutting edges. A sub-ambient pressure is applied to a lumen in the cannula to draw the adipose tissue through the aperture where it is then severed using the cutting edge to disrupt the connective adipose matrix.

In some embodiments of the invention device, optionally in combination with any or all the various embodiments described herein, the needle barrel is between <NUM> and <NUM> gauge in diameter.

In some embodiments of the invention device, optionally in combination with any or all the various embodiments described herein, the needle barrel is between <NUM> and <NUM> in length.

In some embodiments of the invention device, optionally in combination with any or all the various embodiments described herein, the holes contained around the diameter of the needle barrel have a geometry that allows for the cutting and removal of lipoaspirate as the device is pushed forward and backwards within the subcutaneous space.

In some embodiments of the invention device, optionally in combination with any or all the various embodiments described herein, the holes contained around the diameter of the needle barrel have a geometry that allows for the cutting and removal of lipoaspirate as the device is rotated within the subcutaneous space.

In some embodiments of the invention device, optionally in combination with any or all the various embodiments described herein, the device further comprises a guide that prevents the over insertion and under insertion and thus allows collection of subcutaneous adipose tissue at desired depth.

In some embodiments of the invention device, optionally in combination with any or all the various embodiments described herein, the guide is curved to allow puncture of the needle and forces the needle to remain in the subcutaneous layer of adipose tissue.

In some embodiments of the invention device, optionally in combination with any or all the various embodiments described herein, the guide further contains a mechanism that pinches the exterior of the skin and thus raises the subcutaneous fat layer to allow for insertion of the needle.

In some embodiments of the invention device, optionally in combination with any or all the various embodiments described herein, the guide further contains a vacuum mechanism capable of raising the subcutaneous fat layer to allow for insertion of the needle.

In some embodiments of the invention device, optionally in combination with any or all the various embodiments described herein, the device further comprises a blunt cannula where the cannula removably connects with the needle hub.

In another aspect of the present invention, it is provided an adipose tissue collection and purification device as defined in claim <NUM>.

In some embodiments of the invention device, optionally in combination with any or all the various embodiments described herein, the inner syringe contained within the outer syringe is able to evacuate the waste product from lipoaspirated tissue into its barrel and remain contained within its barrel during the reinjection of remaining adipose contained within the filter of the outer syringe.

In some embodiments of the invention device, optionally in combination with any or all the various embodiments described herein, the inner syringe plunger contains a one-way valve that allows for the passage of lipoaspirate waste into its barrel but does not allow the lipoaspirate waste to re-enter the barrel of the outer syringe.

In some embodiments of the invention device, optionally in combination with any or all the various embodiments described herein, the filter is able to contain adipose tissue but is also collapsible thus allowing the adipose to exit the filter and be pushed outside the outer syringe.

In some embodiments of the invention device, optionally in combination with any or all the various embodiments described herein, the inner syringe contained within the outer syringe is able to evacuate the waste product from lipoaspirated tissue into a separate waste product chamber.

In some embodiments of the invention device, optionally in combination with any or all the various embodiments described herein, the filter is a multiple stage filter comprising a plurality of filters where a first stage of filtration uses a filter up to <NUM> micro meters.

In a further aspect of the present invention, it is provided a system for closed adipose tissue harvesting, purficiation, and grafting, defined in claim <NUM>.

In some embodiments of the invention system, optionally in combination with any or all the various embodiments described herein, the needle hub comprises a plurality of needles each of which contains between <NUM>-<NUM> holes around the diameter of the needle barrel which allows passage of adipose tissue and other lipoaspirate.

In some embodiments of the invention system, optionally in combination with any or all the various embodiments described herein, the needle barrel is between <NUM> and <NUM> gauge in diameter.

In some embodiments of the invention system, optionally in combination with any or all the various embodiments described herein, the needle barrel is between <NUM> and <NUM> in length.

In some embodiments of the invention system, optionally in combination with any or all the various embodiments described herein, the holes contained around the diameter of the needle barrel have a geometry that allows for the cutting and removal of lipoaspirate as the device is pushed forward and backwards within the subcutaneous space.

In some embodiments of the invention system, optionally in combination with any or all the various embodiments described herein, the holes contained around the diameter of the needle barrel have a geometry that allows for the cutting and removal of lipoaspirate as the device is rotated within the subcutaneous space.

In some embodiments of the invention system, optionally in combination with any or all the various embodiments described herein, the tissue removal device further comprises a guide that prevents the over insertion and under insertion and thus allows collection of subcutaneous adipose tissue at desired depth.

In some embodiments of the invention system, optionally in combination with any or all the various embodiments described herein, the guide is curved to allow puncture of the needle and forces the needle to remain in the subcutaneous layer of adipose tissue.

In some embodiments of the invention system, optionally in combination with any or all the various embodiments described herein, the guide further contains a mechanism that pinches the exterior of the skin and thus raises the subcutaneous fat layer to allow for insertion of the needle.

In some embodiments of the invention system, optionally in combination with any or all the various embodiments described herein, the guide further contains a vacuum mechanism capable of raising the subcutaneous fat layer to allow for insertion of the needle.

In some embodiments of the invention system, optionally in combination with any or all the various embodiments described herein, the tissue removal device further comprises a blunt cannula where the cannula removably connects with the needle hub.

In some embodiments of the invention system, optionally in combination with any or all the various embodiments described herein, the inner syringe contained within the outer syringe is able to evacuate the waste product from lipoaspirated tissue into its barrel and remain contained within its barrel during the reinjection of remaining adipose contained within the filter of the outer syringe.

In some embodiments of the invention system, optionally in combination with any or all the various embodiments described herein, the inner syringe plunger contains a one-way valve that allows for the passage of lipoaspirate waste into its barrel but does not allow the lipoaspirate waste to re-enter the barrel of the outer syringe.

In some embodiments of the invention system, optionally in combination with any or all the various embodiments described herein, the filter is able to contain adipose tissue but is also collapsible thus allowing the adipose to exit the filter and be pushed outside the outer syringe.

In some embodiments of the invention system, optionally in combination with any or all the various embodiments described herein, the inner syringe contained within the outer syringe is able to evacuate the waste product from lipoaspirated tissue into a separate waste product chamber.

In some embodiments of the invention system, optionally in combination with any or all the various embodiments described herein, the filter is a multiple stage filter comprising a plurality of filters where a first stage of filtration uses a filter up to <NUM> micro meters.

It is also provided a method (not claimed) of using the tissue removal device in a cosmetic application on a subject. For the sake of concise description, the embodiments of the tissue removal device, the tissue collection and purification device, and the system described above are omitted.

In some embodiments of the invention method, optionally in combination with any or all the various embodiments described herein, the subject is a human being.

As used herein, the term "adipose tissue" is used interchangeably with the term "fat", the meaning of which is well known to a person of ordinary skill in the art.

As used herein, the term "adipose tissue removal" or "tissue removal" or "harvesting" or "liposuction" are used interchangeably to mean remove an amount of adipose tissue from a live subject such as a male or female patient.

As used herein, the term "collapsible" refers to the attribute of a material capable of collapsing under pressure or vacuum or capable of changing of shape or contour or of deformation in response to pressure change, and as such, in some embodiments, the term "collapsible" can mean deformable. An example of a material that is collapsible is a plastic or polymeric material forming a bag, e.g., a bag that is described in <CIT> now published as <CIT>.

As used herein, the term "filter" refers to a porous material having a size or size distribution useful for adipose tissue filtration. Some examples of filter useful for the present invention are described in <CIT> now published as <CIT>.

In one aspect of the present invention, it is provided a tissue removal device that allows for the removal of adipose tissue with a needle(s), comprising a needle hub that comprises at least one needle that contains between <NUM>-<NUM> holes around the diameter of the needle barrel which allows passage of adipose tissue and other lipoaspirate.

In some embodiments of the invention device, optionally in combination with any or all the various embodiments disclosed herein, the needle hub comprises a plurality of needles each of which contains between <NUM>-<NUM> holes around the diameter of the needle barrel which allows passage of adipose tissue and other lipoaspirate.

In some embodiments of the invention device, optionally in combination with any or all the various embodiments disclosed herein, the needle barrel is between <NUM> and <NUM> gauge in diameter.

In some embodiments of the invention device, optionally in combination with any or all the various embodiments disclosed herein, the needle barrel is between <NUM> and <NUM> in length.

In some embodiments of the invention device, optionally in combination with any or all the various embodiments disclosed herein, the holes contained around the diameter of the needle barrel have a geometry that allows for the cutting and removal of lipoaspirate as the device is pushed forward and backwards within the subcutaneous space. In some embodiments, the holes contained around the diameter circumference of the needle barrel have a geometry that allows for the cutting and removal of lipoaspirate as the device is rotated within the subcutaneous space.

In some embodiments of the invention device, optionally in combination with any or all the various embodiments disclosed herein, the device further comprising a guide that prevents the over insertion and under insertion and thus allows collection of subcutaneous adipose tissue at desired depth.

In some embodiments of the invention device, optionally in combination with any or all the various embodiments disclosed herein, the guide is curved to allow puncture of the needle and forces the needle to remain in the subcutaneous layer of adipose tissue.

In some embodiments of the invention device, optionally in combination with any or all the various embodiments disclosed herein, the guide further contains a mechanism that pinches the exterior of the skin and thus raises the subcutaneous fat layer to allow for insertion of the needle.

In a further aspect fo the present invention, it is provided an adipose tissue collection and purification device comprising a plurality of self contained syringes wherein each of the syringes comprises an inner syringe included within an outer syringe and wherein a filter is attached inside the outer syringe barrel, the filter having a mesh pore size between <NUM> micro meters and <NUM> micro meters;.

In some embodiments of the invention device, optionally in combination with any or all the various embodiments disclosed herein, the inner syringe contained within the outer syringe is able to evacuate the waste product from lipoaspirated tissue into its barrel and remain contained within its barrel during the reinjection of remaining adipose contained within the filter of the outer syringe.

In some embodiments of the invention device, optionally in combination with any or all the various embodiments disclosed herein, the inner syringe plunger contains a one-way valve that allows for the passage of lipoaspirate waste into its barrel but does not allow the lipoaspirate waste to re-enter the barrel of the outer syringe.

In some embodiments of the invention device, optionally in combination with any or all the various embodiments disclosed herein, the filter is able to contain adipose tissue but is also collapsible thus allowing the adipose to exit the filter and be pushed outside the outer syringe.

An embodiment of the invention device is described in <FIG> (PUREGRAFT <NUM> SYRINGE ASSEMBLY). Referring to <FIG>, element <NUM> is an outer syringe barrel, <NUM> is a piston, <NUM> is a locking pin, <NUM> is a valve (e. g, a check valve), <NUM> is an inner syringe, which is also referred as a female syringe, <NUM> is a filter component, which can be, for example, a polyester mesh (e.g., <NUM> micron with <NUM>% open area), <NUM> is an additional filter component, which can be, for example, <NUM> PPI polyester foam, <NUM> is a further additional filter component, which can be, for example, <NUM> PPI polyester foam, <NUM>-<NUM> comprising a multiple stage depth filter element of the invention device, and <NUM> is an O-ring, which can have a size of [<NUM> X <NUM> ID], made of silicone, e.g. <FIG> shows the assembly of the invention device of <FIG>.

Another embodiment of the invention device is desicribed in <FIG>. Referring to <FIG>, the invention device embodiment includes <NUM>, an outer syringe barrel, <NUM>, a valve, <NUM>, an inner syringe, <NUM>, a filter component (a mesh), <NUM>, an additional filter component (a foam), and a further additional filter component, <NUM> (a <NUM> micron filter disk). In one embodiment, the specifciations of the elements of the device in <FIG> are the same as the corresponding elements of the device of <FIG>.

In a further aspect of the present invention, it is provided a system for closed adipose tissue harvesting, purficiation, and grafting, comrpising an adipose tissue removal component, a collection and purification component, and a grafting component,
wherein:.

<FIG> describes an assembled invention device embodiment of <FIG> in use of tissue harvesting. As shown in <FIG>, a volume of tissue (<NUM>) was collected and cleaned in the outer syringe <NUM>, and the waste (<NUM>) was collected in the inner syringe <NUM>.

In a further aspect of the present invention, it is provided a method of using the device of invention, the adipose tissue collection and purification device of invention and/or the system of invention. In some embodiments, the use includes biomedical and cosmetic applications on a subject. Cosmetic applications can be, for example, organ reshaping or augmentation. Biomedical applications can be, for example, tissue grafting, and cell therapy or tissue regenerative therapies.

Designs of exemplary needles of invention are shown in <FIG>.

<FIG> illustrates an embodiment of the invention wherein a needle hub comprises one or a plurality of needles (<NUM>-<NUM> needles) having a bendable/depressible surface to allow up and down agitation and a method of using the device (Example <NUM>). In some embodiments, the bendable/depressible surface can be an adhesive surface.

A method of using the device of invention of <FIG> can include the following steps:.

To develop a novel and commercially viable method of harvesting small volumes (up to 50cc of adipose tissue, there are three design challenges that must be overcome for this project to be successful. They are as follows:.

Studies on animal models were performed and showed that <NUM>) and <NUM>) requirements were met. The third requirement can be solved using mechanical design of off-the-shelf aseptic ports to transfer the tissue.

The first design requirement focuses on the cannula design. There was a lot of development work done, to determine the correct size and configuration that would work for this application. We know that small needles, <NUM> gauge and up can be insert in the skin little to no pain (28ga. is currently used for insulin injects, OD is <NUM> (. <NUM>") and ID is <NUM> (. For comparison, the smallest cannula used in liposuction procedures has a combined suction opening area of <NUM>. 9x10^-<NUM>^<NUM> (. <NUM> in<NUM>) and we can duplicate that using an array of five of the custom thin wall <NUM>. For our testing we have been testing standard 16ga. needles, as they are easier for me to make custom ground side port openings in my shop. In testing both the 16ga. needle with animal fat, (Pork Bellies) we were able to demonstrate the needles could remove adipose tissue without clogging (See <FIG>). We are working is Vita Needles to make custom cannula needles of the correct size and configuration.

The second design requirement is the ability to capture the adipose tissue and rinse it after it has been harvested. We are currently testing many configuration and material to capture the adipose tissue from the lipoaspirate. Using 3D printer, we have been able to prototype many differing designs and configurations to test to collect the adipose tissue (See <FIG>). We have evaluated differing designs, finding that we can use many different shapes and configurations. Another key to the design is to create fine grid membrane that the tissue will attach to and not be removed with the liposuction or rinsing. We have found one membrane that works well in collecting the adipose tissue. (See <FIG>).

A closed adipose tissue harvesting and cleaning procedure was successfully carried out using an embodiment invention device of <FIG>. As shown in <FIG>, a volume of tissue (<NUM>) was collected and cleaned in the outer syringe <NUM>, and the waste (<NUM>) was collected in the inner syringe <NUM>. The cleaned adipose tissue <NUM> is ready for use.

We have successfully combined these requirements into one device that has been used to harvest animal fat. We tested both needle cannulas with animal fat to show that tissue could be collected without clogging of the cannulas. The first test was with the 16ga needle and the tissue separation device and was able to collect <NUM>-<NUM> grams in just a few minutes. (See <FIG>) Using animal fat (Pork Bellies) is considered a worst case test for us, in that we cannot really introduce tumescent fluid, and the tissue is at room temperature. But that we were able to collect measurable amount of tissue in a short time was significant. Our initial goal is to be able to collect <NUM> to <NUM> grams of adipose tissue in less than <NUM> minutes. We tested using a 22ga. needle modified with <NUM> side ports. Again, we were able to collect measurable amount of animal tissue in a short period of time. (See <FIG>) Comparing the two images, you can see that the 22ga. needle is much smaller, but able to get about the same amount of adipose tissue.

Claim 1:
A device for the removal of adipose tissue with needles configured to remove the adipose tissue, the device comprising a needle hub (<NUM>) that comprises a plurality of needles (<NUM>) each of which contains between <NUM>-<NUM> holes (14b) around the diameter of a needle barrel which allows passage of adipose tissue and other lipoaspirate.