Double-lumen oocyte aspiration needle and use thereof

Abstract

The present invention relates to a double-lumen oocyte aspiration needle and use thereof in the collection of oocytes from the ovaries of animals undergoing assisted reproduction procedures.

Description

DOUBLE LUMEN OOCYTE ASPIRATION NEEDLE AND ITS USE FIELD OF INVENTION

[0001] The present invention is in the fields of Animal Science, Veterinary Medicine, Biology and Engineering and is intended for a double lumen oocyte aspiration needle and its use in collecting oocytes from animal ovaries in assisted reproduction procedures. BACKGROUND AND FUNDAMENTALS OF THE INVENTION

[0002] Assisted reproduction procedures in animals have become widely accepted and applied because they facilitate or enhance the encounter between male and female gametes so that fertilization can occur, which can be applied to the animal in vivo, as well as in an in vitro laboratory, thus generating better reproductive efficiency and genetic improvement in agricultural practice.

[0003] In the process of aspiration or recovery of oocytes, a needle is introduced which, in contact with the ovary, pierces a follicle in which the oocytes are collected by suction through a hose connected to the end opposite the bevel of the needle and integrated into a container consisting of a cap and a test tube that communicates with a vacuum source. The oocyte, once sucked by the needle cannula, passes through a hose and is subsequently deposited in a test tube.

[0004] The double lumen oocyte aspiration needle consists of an external needle (responsible for piercing the animal's follicle) with an internal cannula (which aspirates the desired material). The space between the diameter of the needle and the internal cannula forms a passage for the application of fluid, used to wash the punctured region. The cannula The internal cannula is connected to a vacuum system for aspiration and the space between the needle and the internal cannula is connected to a second hose in which the fluid is applied through a syringe or bag. This hydraulic system uses a properly sealed Y-type conductor to ensure that the fluid is directed to the desired end (needle tip). In this model, the Y system ends up being very large and, therefore, needs to be outside an aspiration guide (adapter used with ultrasound and passage channel for the needle). Therefore, it is necessary to compensate for the stability of the system by using a longer needle.

[0005] In the state of the art, models of double-lumen oocyte aspiration needles are known, which consist of a long needle connected to a two-way chamber at the end opposite the needle bevel, and in which one of the ways contains a hose where fluids are applied for irrigation, and in the other way a cannula with a diameter relatively smaller than the internal diameter of the needle, which runs along its entire length until close to the needle bevel. This cannula is also connected to a hose, at its end opposite the needle bevel, which leads to the container composed of a cap and a test tube. A seal is necessary in this two-way chamber to prevent fluid leakage and to ensure that the oocytes follow the process flow. Thus, in the oocyte aspiration or recovery procedure, the injected fluid helps to detach the oocytes from the follicles and promotes better fluidity in the aspiration.To have a better constructive understanding, a double lumen oocyte aspiration needle basically comprises a smaller channel (cannula. (internal) within a larger channel (needle lumen). A fluid is injected into the space created between both channels, aiding in the aspiration of oocytes through the smaller channel (internal cannula). The two-way chamber connected to the opposite end of the needle bevel is usually "Y" or "T" shaped and has a hose connection for fluid application and a hose connection that is directly connected to the internal cannula, isolated from the entire chamber that directs the fluid to the space between the channels. In this way, two pathways are provided, one for injection and the other for aspiration, in which the aspirated fluid is expelled at the end close to the bevel and then aspirated through the internal cannula along with the material to be collected.

[0006] In order to perform this procedure of collecting or recovering oocytes by aspiration in large animals such as horses and cattle, it is necessary to visually monitor the process by ultrasound, which is performed intravaginally in the animal by means of an adapter device (aspiration guide), which accommodates the transducer and protects the entire system with the needle in a cavity in the device itself, thus making it possible to introduce the needle and transducer through the vaginal canal without causing injury to the animal. In general, these adapter devices (aspiration guides) are made in an anatomical way and with the smallest possible dimensions, in order to avoid discomfort for the animal, resulting in a cavity with limited dimensions for accommodating the needle in said device.

[0007] International publication WO 93 / 07917 refers to a double lumen oocyte collection device. The referred The oocyte collection device includes parallel cannulas that extend axially outward from a proximal end of the connection / hub. The device also provides a finger depression on the connection / hub. Such a depression located on the connection / hub directly corresponds to an elongated beveled point of a cannula that extends from the distal end of the connection / hub. The unique parallel cannulas and finger depression on the connection / hub allow medical professionals to uniformly rotate and precisely orient the beveled tip of the device to puncture a follicle in an ovary with minimal trauma prior to oocyte collection.

[0008] International publication WO 2009 / 040493 relates to a sampling needle, in particular a needle suitable for obtaining oocytes from humans and animals, the needle comprising a first tubular region in fluid communication with a second tubular region, the first region comprising a guide terminal for insertion into a human or animal and a second tubular region comprising a movable terminal region for fluid communication, in terms of receiving a fluid, the first tubular region having an external diameter that is smaller than the internal diameter of the secondary tubular region. The said prior art document also relates to a method of obtaining a sample, in particular a human or animal oocyte.

[0009] The US patent application US 2008 / 091119 Al refers to a device and method for verifying the characteristics of follicular fluid in vivo, comprising an aspiration needle for obtaining oocytes having one or more sensors arranged in the external cannula or incorporated into its outer cannula wall for sensing and measuring characteristics of the constituents of the follicular fluid, such as pH level and oxygen concentration. The sensors are micro sensors that communicate via physical connection or wireless connection with a meter and display. The measurements are recorded and, optionally, printed for future review and selection of the most viable oocytes. The method involves inserting an aspiration needle into the ovary with one or more sensors. The needle is guided to a first follicle and inserted into a first follicle. The characteristics of the follicular fluid are monitored and measured as visualized on the meter and recorded for future reference and selection.

[0010] International publication WO 2010 / 054660 Al refers to a dual cannula system for oocyte retrieval. The dual cannula system has an inner cannula and an outer cannula, with the inner cannula operating freely within the outer cannula. In addition, the inner cannula can be removed from the outer cannula, and both cannulae have a tip that is pointed and beveled. The tip of the inner cannula of the dual cannula system is in front when introduced into a subject, and the inner cannula is used to locate one or more anesthetic parts when the inner cannula is procedurally introduced into a subject. The tip of the outer cannula runs behind the tip of the inner cannula. The inner cannula is removed before cells and / or body fluid are obtained through the outer cannula.

[0011] It is known that, in most models of double-lumen oocyte aspiration needles, a typical construction consists of a long needle of approximately 500 mm, for movement within the cavity of these adapter devices (aspiration guides), with an internal cannula joined in a chamber, which directs the fluid between the gap generated by a gap in the cannula and the internal part of the needle, which also promotes a seal on the end of the internal cannula where a hose is attached to collect the aspirated material.

[0012] Due to the anatomy of large animals, these adapter devices (aspiration guides) are long, which requires a needle with a length of around 500 mm, causing a negative effect on the act of aspiration, as it subjects the oocyte to a long journey through a metal area with a high coefficient of friction, which causes wear on these cells.

[0013] Planer Limited markets a double lumen needle, called "Kitazato", which allows oocytes to be aspirated and follicles to be released in the same procedure, through different channels. The double lumen needle has two channels inside, with a central channel of larger diameter that allows the aspiration of oocytes and follicular fluid. There is also a second channel that allows the release of the sterilizing agent for cleaning the follicle. The double channel needle marketed by this company is also a needle whose construction requires a long needle cannula and internal metal tubing.

[0014] The company Vitroli fe Group markets the product called "Sense™ Single and Double Lumen", which has a 16G needle body and a 19G needle tip. This is a double channel needle that stands out for the staggered length of the needle cannula, which reduces the diameter in the bevel region in order to provide a smaller area in diameter in the perforated region, but always following the conventional construction of the state of the art of long conventional double flow needles.

[0015] Cook Medical® markets a product called "Cook® EchoTip® Double Lumen Aspiration Needle", which is used to aspirate and release oocytes from ovarian follicles. This is a conventional double lumen needle, with the advantage of having micro markings in the bevel region that better reflect ultrasound, thus improving the visualization and location of the needle during the aspiration process.

[0016] It is also evident, from the state of the art, that the technical problem yet to be solved is the reduction of lesions and loss of oocytes in quality and / or viability, due to the friction of the oocytes on the internal walls of the tubing of the aspiration devices.

[0017] Taking into account that aspiration needles are single-use and disposable items, devices containing long needles with average measurements of 500 mm in length and with an internal metal tube become a considerable problem in the disposal of these materials, directly impacting the issue of generation of waste to be later incinerated, also impacting the environmental issue.

[0018] In view of all of the above, the Applicant has developed a double-lumen oocyte aspiration needle that has a compact construction configuration, featuring a bifurcated chamber, ensuring a watertight seal of the hydraulic system, as well as a needle for reduced size aspiration, promoting the aspiration of a greater quantity and quality of viable oocytes, since the aspirated oocytes do not have any type of contact with the metal parts of the device with a high coefficient of friction. DESCRIPTION OF FIGURES

[0019] Figure 1 illustrates a perspective view of the compact construction configuration of the double lumen oocyte aspiration needle of the present invention.

[0020] Figure 2 illustrates a lateral sectional representation of the compact construction configuration of the double lumen oocyte aspiration needle of the invention.

[0021] Figure 3 illustrates a lateral representation of an enlarged sectional detail of the compact construction configuration of the invention.

[0022] Figure 4 illustrates an exploded view of the components in rear perspective of the needle of the present invention.

[0023] Figure 5 illustrates a lateral sectional representation of the rear base component of the compact construction configuration of the present invention.

[0024] Figure 6 illustrates a lateral sectional representation of the front base component of the compact construction configuration of the present invention.

[0025] Figure 7 illustrates a lateral sectional representation of the compact construction configuration of the double lumen oocyte aspiration needle of the present invention mounted on an adapter device (aspiration guide).

[0026] Figure 8 illustrates a linear drawing representation of a veterinarian performing oocyte aspiration using the double lumen aspiration needle of the present invention. invention .

[0027] Figure 9 shows a photograph taken on a Nikon SMZ 800N stereoscope using a PrimeCam INTERVIS ION SMZ 12 camera, which shows the comparison between oocytes aspirated with a double-lumen oocyte aspiration needle of the present invention in group A and aspirated with a double-lumen aspiration needle with a 500 mm long cannula, of the state of the art, in group B. DETAILED DESCRIPTION OF THE INVENTION

[0028] In some alternatives for double-lumen oocyte aspiration needles, a reduced bifurcated chamber would be provided in order to accommodate the entire assembly within the cavity of these adapter devices (aspiration guides), thus enabling the reduction of the length of the needle and the internal aspiration cannula, which would provide the oocyte with less friction on its cells, due to the smaller area of ​​contact with the metal cannula until it reaches the hose, which is normally made of polymers with a low coefficient of friction. These needle models should be used with the aid of a rod screwed or glued to this bifurcated chamber, necessary for its movement when piercing the follicles.However, one of the main problems found in double lumen oocyte aspiration needles with a reduced bifurcated chamber is the construction difficulty, since it is a system that requires a good seal, due to the pressure applied when injecting the fluid. This seal would be made by some type of glue that, hypothetically, would not guarantee good repeatability in production and would generate leaks in the system, not guaranteeing a good flow of fluid for washing the fluid. follicle. Another option would be to manufacture by overinjection of thermoplastic over the internal metal cannula and glued or screwed into the bifurcated chamber, which would make the production process with good repeatability, but with a higher cost, remaining the problem of the metal cannula with the high coefficient of friction.

[0029] The present invention relates to a double lumen oocyte aspiration needle and its use in collecting oocytes from animal ovaries in assisted reproduction procedures.

[0030] In a first embodiment, the patent application provides a double-lumen oocyte aspiration needle that has a bifurcated chamber, ensuring a watertight seal of the hydraulic system, as well as a 12G aspiration needle (gauges), 3 mm in diameter and with a total length of less than 110 mm, on average 5 times smaller than traditional state-of-the-art needles, through which a low friction coefficient tubing passes internally, promoting the aspiration of a greater quantity and quality of viable oocytes, since the aspirated oocytes do not have any type of contact with the metal parts of said device.

[0031] The compact construction configuration of the aspiration needle revealed by the present invention, due to its tightness, allows to avoid leaks and loss of pressure during aspiration, which directly affect the obtaining of viable oocytes in animals.

[0032] The present invention also proposes the replacement of the internal channel (metal cannula used in the state of the art) by a hose in thermoplastic material. with a low coefficient of static friction of approximately 0.05 p, which is fixed and isolated, by compression by means of a silicone rubber sealing tube, together with an adhesion material, to the entire internal watertight hydraulic system composed of the bifurcated chamber and the reduced-size needle. Such sealing tube accommodated in the cavity properly designed to create sufficient tightness around said hose in order to fix it, also creates a watertight seal of the bifurcated chamber composed of the front base piece and the rear base piece, generating better direction for the fluid injected in the follicle washes.

[0033] Furthermore, the present invention provides a double-lumen oocyte aspiration needle that, in addition to being compact, is completely disposable, without the need for assembling parts in the field, which also reduces contamination rates when producing embryos from animals such as cattle, horses, pigs, among others.

[0034] The compact construction configuration of the double-lumen oocyte aspiration needle of the present invention consists of a suction hose (4), made of a thermoplastic material with a low friction coefficient, inserted internally throughout the entire sealed hydraulic system, that is, from the internal space of the needle (1) connected to a bifurcated chamber composed of the front base piece (2) which, in turn, is coupled to the rear base piece (3) to a rod (19) which acts as an extension for moving the compact construction configuration of the double-lumen oocyte aspiration needle within an "aspiration guide" adapter device (21).

[0035] In general, the constructive configuration The compact double-lumen oocyte aspiration needle of the present invention comprises a needle (1) fitted to the duet (17) and joined in the attachment region (7) with adhesion material, in particular epoxy glue, wrapped around the housing (18) internal to the front base piece (2), whereby the duet (17) limits the depth of the needle (1) to be fitted. The housing (18) is a concave profile that prevents the detachment of the attachment region (7) with adhesion material, in particular epoxy glue, after solidification.

[0036] In the rear base part (3), the suction hose (4) is passed through the channel (14), being kept pressed by the silicone rubber sealing tube (6) housed in a subsequent cavity (15) and aligned with the channel (14). The suction hose (4) is slightly curved in a relief and communication space (8), through which it is routed internally to the needle (1) until close to the bevel (11), leaving a space (9) between the internal walls of the needle (1) and the external surface of the suction hose (4).

[0037] The relief and communication space (8) creates a direct communication between the spacing (9) and the communication channel (13) which is directly connected to the irrigation inlet duct (10), which is then connected to the irrigation hose (5).

[0038] The front base piece (2) fits, in a watertight manner, into the cavity (16) in the rear base piece (3), which has a thread (12) used to couple the entire hydraulic system to a rod (19) that acts as an extension for moving the compact construction arrangement in a double lumen suction needle within a "suction guide" adapter device (21).

[0039] In particular, the compact construction configuration of the needle of the present invention allows the insertion of the entire assembly within the suction guide, in addition to not using metal tubes internally.

[0040] In particular, the needles used by the present invention have an approximate external diameter of 3 mm, corresponding to the 12G needle type with a length of 110 mm.

[0041] In a second embodiment, the present patent application deals with the use of the compact construction configuration of the aspiration needle of the present invention in the collection of oocytes from animal ovaries, in an assisted reproduction procedure.

[0042] In general, during the oocyte collection and recovery procedure, ultrasound monitoring is used to guide the needle cannula to the correct position in the ovary. The needle bevel pierces the follicle and, to facilitate the detachment and drainage of the oocytes, fluids are injected through the irrigation hose, passing through the irrigation inlet duct, which continues through the communication channel, until it reaches the relief and communication space, where the injected fluid fills the entire space and begins to move through the space until it exits through the needle cannula. At this moment, the silicone rubber sealing tube, which is pressed between the cavity and the suction hose, acts as a seal, preventing the injected fluid from returning through the duct.

[0043] In continuity, the suction hose connected to a lid on a test tube, also connected with the same lid to a vacuum generation source (24) will perform the suction of the oocytes along with the fluid that is injected.

[0044] The technical advantage of using the compact construction configuration of the double-lumen oocyte aspiration needle of the present invention, in particular, of the application of the aspiration hose made of thermoplastic material with a low coefficient of friction and of the compact and watertight construction configuration, is that such construction reduces by up to 40% the damage caused by friction between the body of the oocyte and the path taken during aspiration, as is the case with double-lumen needles that allow contact with the aspirated oocytes with the metal parts of the needle. Thus, the problem of obtaining viable and healthy oocytes is solved by using the compact construction configuration of the aspiration needle of the present invention.

[0045] Also, the use of the front base piece accommodated with the rear base piece associated with the silicone rubber sealing tube eliminates the need to use long needle cannulas and long metal tubes. DETAILED DESCRIPTION OF THE INVENTION

[0046] According to figure 1 of the present patent application, the compact construction configuration of the needle of the invention is composed of a suction hose (4) and an irrigation hose (5) connected to the bifurcated chamber, composed of a rear base piece (3) coupled to a front base piece (2), and a needle (1).

[0047] With regard to figure 2, the compact construction configuration of the double lumen suction needle demonstrates the entire length of the suction hose (4), along the sealed hydraulic system, passing through the needle (1), front base piece (2), relief and communication space. (8) , silicone rubber sealing tube (6), communication channel (13) , rear base piece (3) , coupling thread (12) , irrigation inlet duct (10) and irrigation hose (5) .

[0048] In figure 3, all the details of the internal sealed hydraulic system of the compact construction configuration of the double lumen suction needle of the present invention are shown, highlighting the arrangement of the bifurcated chamber, which is composed of the connection of the rear base part (3) coupled to a front base part (2) and its sealing parts, for example, the fixing region (7) with adhesion material, in particular, epoxy glue and the sealing tube (6) made of silicone rubber.

[0049] Figure 9 refers to a photograph obtained by a Nikon SMZ800N model stereoscope with a zoom ratio of 8:1 (zoom range of lx-8x) and a resolution of 640LP / mm (using a Plan ED Apo 2x / WE objective at maximum zoom) with its high numerical aperture optics. Semi-apochromatic optics through a high-resolution 4K digital camera from PrimeCam model INTERVISION SMZ12, 12MP connected via cable to a computer with a Windows-based operating system, for example, Windows 10, which presents the results of a comparative experiment on the quality of aspiration of oocytes obtained, carried out between a conventional double-lumen needle with a metal cannula measuring more than 500 mm (B) and with the compact construction configuration of the needle of the invention (A) to verify the quality of viable oocytes.It is clearly noted in figure 9, according to experimental results in (B), the excessive wear of the cell walls of the oocytes aspirated by the conventional needle and the preservation of the oocytes aspirated by. compact constructional configuration of the double lumen aspiration needle of the present invention, as indicated in the experimental results presented in (A).

[0050] Initially, the recovery of oocytes from four healthy Quarter Horse mares, from homogeneous genetic groups, with similar ages of 3 years, with the following weights, was evaluated: 512 kg for mare A, 533 kg for mare B, 522 kg for mare C and 499 for mare D.

[0051] In mares A and B, the 12G double-lumen aspiration needle of the present invention was used, while in mares C and D, the conventional 12G long double-lumen needle was used. Both were aspirated using a model WTA-BV005d vacuum pump and the oocyte washing fluid used was Dulbecco's modified phosphate buffer saline. - DMPBS of the Reprodux brand and whose composition is calcium chloride dihydrate - 0.133 g / l, magnesium chloride hexahydrate - 0.100g / l, potassium chloride - 0.200g / l, monobasic potassium phosphate - 0.200g / l, sodium chloride 8.000 g / l, dibasic sodium phosphate - 1.150 g / 1, glucose - 1.000 g / 1 and gentamicin - 0.050 g / 1.

[0052] Throughout the entire aspiration process, the pressure used by all needles was between 120 mmHg and a maximum of 150 mmHg with a controlled temperature for the collection tube at 39°C.

[0053] After collecting the oocytes, the aspirated material was transferred to a filter, where it was washed with the same solution used in the aspiration, and separated into two groups in a Petri dish for observation and analysis in a Nikon SMZ800N stereoscope with a zoom ratio of 8:1. (lx-8x zoom range) and 640LP / mm resolution (using Plan ED Apo 2x / WE objective at maximum zoom) with its high numerical aperture optics. Semi-apochromatic optics through a 4K high-resolution digital camera from PrimeCam model INTERVISION SMZ12, 12MP connected via cable to a computer with Windows 10. As shown in Figure 9, the aspirated oocytes (right side) and identified as (B) were aspirated with a conventional 12G double-lumen needle and metal cannula measuring more than 500 mm and the aspirated oocytes (left side) and identified as (A) were aspirated with the 12G double-lumen aspiration needle of the present invention, where it is clearly possible to observe the cell wear that occurred in the oocytes on the right, due to friction on the inner walls of the cannula of the conventional long needles, while on the left side, we can notice greater preservation of the cells. METHODOLOGY FOR FOLLICULAR ASPIRATION

[0054] Preparation of donor and recipient mares: The donor mares were selected based on their high genetic potential, even if they cannot conceive naturally, the recipient mares are healthy and fertile females. Hormonal stimulation: The veterinarian specialized in equine reproduction performed hormonal stimulation on the donor and recipient mares through the application of progesterone, this stimulation aims to develop multiple ovarian follicles in the donor mares, where the oocytes are found. Use of ultrasound: The follicular aspiration procedure requires the use of ultrasound, the veterinarian used a modern Mindray ultrasound device with a probe (transducer) model 65EC10EB adapted to a Sealed WTA aspiration guide with an 8 mm mandrel for visualizing ovarian follicles in donor mares. Follicular aspiration procedure: With the mare sedated and anesthetized, the veterinarian introduced the aspiration guide connected to an aspirator (vacuum pump model BV005 from WTA) through the vagina parallel to the cervix. Using the images obtained by ultrasound, the professional was able to guide the aspiration guide to the ovarian follicle containing the oocyte. An aspiration needle was aligned with the follicle and the oocyte was carefully aspirated. The procedures were performed on one or more follicles per individual, depending on the response to hormonal stimulation. Oocyte processing: the collected oocytes were washed, separated and then processed in the laboratory to be analyzed and compared and analyzed for their cellular preservation.

Claims

CLAIMS 1. DOUBLE LUMEN OOCYTE ASPIRATION NEEDLE, characterized by comprising a needle (1) connected to a bifurcated chamber composed of a front base piece (2) and coupled to a rear base piece (3) and a shaft (19); in which said needle (1) has an aspiration hose (4) inserted internally to said needle (1); in which said aspiration hose (4) is made of a thermoplastic material with a low coefficient of friction; and in which said needle (1) has a total length of less than 110 mm.

2. NEEDLE, according to claim 1, characterized in that said thermoplastic material of the suction hose (4) has a static friction coefficient of 0.05 p.

3. NEEDLE, according to claim 1, characterized in that said suction hose (4) is fixed and isolated, by means of a silicone rubber sealing tube (6), together with an adhesion material, to the entire internal sealed hydraulic system composed of the bifurcated chamber and said needle (1).

4. NEEDLE, according to claim 3, characterized in that said adhesion material comprises an epoxy glue.

5. NEEDLE, according to claim 1, characterized in that said needle (1) is fitted to a duet (17) and joined in the fixing region (7) with adhesion material wrapped around the housing (18) inside said front base piece (2).

6. NEEDLE, according to claim 5, characterized in that said adhesion material comprises an epoxy glue.

7. NEEDLE, according to claim 1, characterized in that said suction hose (4) is passed through the channel (14) and kept pressed by the silicone rubber sealing tube (6) housed in a subsequent cavity (15) and aligned with the channel (14).

8. NEEDLE, according to any one of claims 1 to 3, characterized in that said suction hose (4) is slightly curved in a relief and communication space (8), through which it is routed internally to said needle (1) until close to the bevel (11), leaving a space (9) between the internal walls of said needle (1) and the external surface of said suction hose (4).

9. NEEDLE, according to claim 8, characterized in that said relief and communication space (8) creates a direct communication between said spacing (9) and the communication channel (13) that is directly connected to the irrigation inlet duct (10), which remains connected to the irrigation hose (5).

10. NEEDLE, according to claim 1, characterized in that the front base part (2) fits tightly into the cavity (16) in the rear base part (3), said rear base part (3) having a thread (12) for coupling the hydraulic system to a rod (19).

11. USE OF THE DOUBLE LUMEN OOCYTE ASPIRATION NEEDLE, as defined in any one of claims 1 to 10, characterized in the collection of oocytes from animal ovaries, in assisted reproduction procedures.

12. USE, according to claim 11, characterized in that the animals include cattle, horses and / or pigs.

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