EXTRACORPOREAL BIOENGINEERED DUAL-CELL LIVER REGENERATION SYSTEM (EBDLR) AND BIO PURIFIER THEREFOR

An EBDLR system includes a multi-layered bio purifier having a plurality of layers. Each layer includes a membrane, a first type of cells on a first side of the membrane in a first channel, and a second type of cells on a second side of the membrane in a second channel. The EBDLR includes a plasma separator to receive blood from a subject and separate a plasma component from the blood, a first reservoir to collect the plasma component, and a second pump to move the plasma component from the first reservoir to the multi-layered bio purifier. The multi-layered bio purifier distributes the plasma component into the first and second channels of each layer to purify the plasma component. The EBDLR includes a second reservoir to collect the purified plasma component and a third pump to infuse the purified plasma component from the second reservoir into the subject.

TECHNICAL FIELD

The present disclosure relates to medical devices, and more particularly to methods, techniques, and systems for treating a subject with acute liver failure using an extracorporeal bioengineered dual-cell liver regeneration system (EBDLR).

BACKGROUND

Liver is one of the most vital organs of the human body. It plays a fundamental role in metabolic activities such as the regulation of glucose levels in the blood, production of plasma proteins, drug metabolism, production of bile, and presents a complex structure from a morphological and functional point of view. Being so indispensable, this organ is also prone to injury and damage. Although, the liver is known to have excellent regeneration capacity, under certain conditions, it fails to recover. Acute liver failure is one such condition which is classified as a medical emergency. Acute liver failure is a medical condition where the liver stops functioning in a short span of time. Orthotropic liver transplant is currently the most sought out treatment modality across the globe. With a success rate of 75%, it is often presented with challenges where 50% of the patients may not receive a liver transplant.

The drawings described herein are for illustrative purposes and are not intended to limit the scope of the present subject matter in any way.

DETAILED DESCRIPTION

Acute liver failure (ALF) is a life-threatening illness, where a normal liver fails within days to weeks. Sudden loss of synthetic and detoxification function of liver results in jaundice, encephalopathy, coagulopathy, and multiorgan failure. The etiology of acute liver failure varies demographically. In some places, acute viral hepatitis is the most common cause of acute liver failure. The mortality of acute liver failure is as high as 40-50% and causes of death in acute liver failure include brain herniation due to raised intracranial pressure (35%) and sepsis with multi-organ failure. Acute liver failure occurs when liver cells are damaged significantly and are no longer able to function.

Some existing methods and systems are designed keeping in mind the requirements of managing patients with acute liver failure in clinics/hospitals. The current modality of treatment includes medications to reduce liver damage and antidotes against the causative agent. For example, N-acetylcysteine and activated charcoal is routinely administered in cases of the drug-induced acute liver failure. Extremely critical cases usually require emergency liver transplants. The problems associated with the liver transplant are diverse. They range from finding a suitable donor to the costs associated with the surgery. Treating a patient diagnosed with the acute liver failure may be a race against time.

Examples described herein may provide an enhanced method, technique, and system to treat patients suffering from an acute liver failure with a bioengineered solution in the form of an extracorporeal bioengineered dual-cell liver regeneration (EBDLR) system. In an example, the EBDLR system may include a multi-layered bio purifier having a plurality of layers. Each layer may include a membrane, a first type of cells on a first side of the membrane in a first channel, and a second type of cells on a second side of the membrane in a second channel. Further, the EBDLR system may include a plasma separator to receive blood from a subject via a first pump and separate a plasma component from the blood, a first reservoir to collect the plasma component, and a second pump to move the plasma component from the first reservoir to the multi-layered bio purifier at a predefined flow rate. The multi-layered bio purifier may distribute the plasma component into the first channel and the second channel of each layer to purify the plasma component. Further, the EBDLR system may include a second reservoir to collect the purified plasma component from the multi-layered bio purifier and a third pump to infuse the purified plasma component from the second reservoir into the subject.

In some existing example techniques, growing cells (i.e., hepatocytes and endothelial cells) has been employed in the field of lab/liver-on chip models. In this example, a small number of hepatocytes are grown on a microbloodic platform for experiments to test drug toxicity. The cell number in such applications is usually in the range of 102-103. In the examples described herein, the EBDLR system uses an exponentially higher number of cells (e.g., 108-109). Even though the process of scaling the cell number from 103to 109may appear linear, there are complex parameters and calculations involved. The non-linear nature of scaleup associated with biological components like cells is well known. Further, the plasma flow associated with these cells is maintained at a critical rate in the range of 100-500 μL/min. This flow rate technically is classified as a creeping flow. Biological bloods like blood/plasma flowing though capillary is a classic example of a naturally occurring creeping flow. The drag associated with creeping flows is usually high. Thus, initiating and maintaining a creeping flow rate is a difficult condition. Examples described herein may provide a constant creepy flow in the bioreactor/bio purifier. This has been achieved by critically estimating the requirements for the first and second channel dimensions (e.g., serpentine channel dimensions).

Further, the bio purifier described herein may use 2 cell types in a specialised arrangement which allows the cells to crosstalk. The 2 types of cells are hepatocytes and endothelial cells. The arrangement to create a co-culture is derived based on inspiration from the basic understanding of human physiology. In the examples described herein, the EBDLR system, which is an external device, includes full functioning capacity of the liver. The EBDLR system is a lightweight and portable device that can be connected to the host suffering from the acute liver failure. Impure blood from the host is pumped through the device which detoxifies the blood and sends it back into the patient.

Thus, the EBDLR system is built to provide external hepatic support which can be run on a continues basis. The EBDLR system may maintain the hemodynamic parameters of the patient during the procedure. For example, blood withdrawal from the patient may results in peripheral blood pressure drop. This can also lead to the collapse of blood vessels connected to the EBDLR system. Hemodynamic instability is often reported in patients undergoing renal dialysis. In the examples described herein, the flow rates at which the EBDLR system operates is extremely safe (0-300 mL/min). This flow rate does not exert excessive shear stress on the blood cells therefore, it prevents haemolysis and hemodynamic instability. The use of state-of-the-art pressure sensing mechanism makes the EBDLR system one of the safe biomedical devices built for assisting clinicians in managing critically ill patients.

In the following description, for purposes of explanation, numerous specific details are set forth in order to provide a thorough understanding of the present techniques. However, the example apparatuses, devices, and systems, may be practiced without these specific details. Reference in the specification to “an example” or similar language means that a particular feature, structure, or characteristic described may be included in at least that one example but may not be in other examples.

Referring now to the figures,FIG.1Ais a schematic diagram of an example extracorporeal bioengineered dual-cell liver regeneration (EBDLR) system100including a multi-layered bio purifier102. EBDLR system100may refer to a lightweight and portable device that can be connected to a host (e.g., a human, dog, and the like) suffering from an acute liver failure. Impure blood from the host is pumped through EBDLR system100which detoxifies the blood and sends the detoxified blood back into the host. Thus, EBDLR system100may provide an external-hepatic support to treat hosts (e.g., patients) suffering from the acute liver failure with a bioengineered solution. In this example, EBDLR system100may take over the functions of a native liver (both synthetic and detoxifying) when connected to the host.

EBDLR system100may employ principles of engineering and biotechnology in a sync. The engineering aspect of EBDLR system100takes care of a safe method to withdraw and circulate blood and plasma through EBDLR system100. Biotechnology approach of EBDLR system100ensures that the plasma is detoxified by specialised cells (e.g., hepatocytes and endothelial cells) present in multi-layered bio purifier102. Thus, external detoxification using EBDLR system100may enable the native/injured liver to regenerate aiding the patient's survival.

EBDLR system100is a state-of-the-art ensemble of peristaltic pumps, pressure transducers, flow sensors, clamps, and the like. The hardware is electronically integrated to a processor which is responsible for efficient and trouble-free functioning of EBDLR system100. The processor may be internal or external, but connected, to EBDLR system100. EBDLR system100systematically draws blood from the patient without affecting the native blood pressure. EBDLR system100may include multi-layered bio purifier102having a plurality of layers104. An example layer of plurality of layers104is explained with respect toFIG.1B.

Referring now toFIG.1B,FIG.1Bis an exploded schematic diagram of an example layer104of multi-layered bio purifier102ofFIG.1A. Each layer104may include a membrane152, a first type of cells154on a first side156of membrane152in a first channel158, and a second type of cells160on a second side162of membrane152in a second channel164. First channel158and second channel164may be defined on a first substrate168and a second substrate176, respectively. The first type of cells154may include hepatocytes and the second type of cells may include endothelial cells. In an example, multi-layered bio purifier102may include a total number of the first type of cells in a range of 106-1010cells and a total amount of the second type of cells in a range of 106-1010cells. The total number of the first type of cells and the second type of cells may be grown in multi-layered bio purifier102may depend on the type of subject.

Further, each layer104may include a first base member166disposed on a first adhesive layer (e.g., first substrate168). First base member166may include inlets170A and170B to receive the plasma component. Further, first base member166may include outlets174A and174B to output the purified plasma component. Furthermore, each layer104may include a second base member172disposed on a second adhesive layer (e.g., second substrate176). First base member166and second base member172may form housing for each layer104. In some examples, inlets170A and170B and outlets174A and174B may be provided in first base member166, second base member172, or a combination thereof. In some examples, inlets170A and170B may be provided on first base member166and outlets174A and174B may be provided on second base member172.

Referring back toFIG.1A, multi-layered bio purifier102may include a first manifold assembly146to split the plasma component into the first channel (e.g.,158) and the second channel (e.g.,164) of the plurality of layers. In an example, first manifold assembly146may include a first manifold layer and a second manifold layer. For example, the plasma component is split into a first portion and a second portion. In this example, the first manifold layer may further split the first portion of the plasma component into a plurality of first paths. Each first path may be connected to an input (e.g., inlet170A) of the first channel of one of the plurality of layers. Further, the second manifold layer may split the second portion of the plasma component into a plurality of second paths. Each second path is connected to an input (e.g., inlet170B) of the second channel of one of the plurality of layers. Each inlet170A and170B may be connected to first manifold assembly146.

Multi-layered bio purifier102may include a second manifold assembly148to combine the purified plasma component from the first channel and the second channel of the plurality of layers. In this example, second manifold assembly148may include a third manifold layer to receive and combine the processed plasma component from a plurality of third paths. Each third path is connected to an output (e.g., outlet174A) of the first channel of one of the plurality of layers. Also, second manifold assembly148may include a fourth manifold layer to receive and combine the processed plasma component from a plurality of fourth paths. Each fourth path may be connected to an output (e.g., outlet174B) of the second channel of one of the plurality of layers. Each outlet174A and174B may be connected to second manifold assembly148.

Further, EBDLR system100may include a thermally regulated chamber138to house multi-layered bio purifier102. Thermally regulated chamber138may maintain a defined temperature of the plasma component passing through multi-layered bio purifier102. In some examples, EBDLR system100may include a first temperature sensor140to sense a temperature of the plasma component entering multi-layered bio purifier102and a second temperature sensor142to sense a temperature of the plasma component leaving multi-layered bio purifier102. Thus, multi-layered bio purifier102is housed in a sophisticated thermally regulated chamber to ensure that the plasma passing through multi-layered bio purifier102does not overheat/cool or get dehydrated.

Further, EBDLR system100may include a plasma separator106to receive blood from a subject (e.g., patient) via a first pump108and separate the plasma component from the blood. In some examples, EBDLR system100may include an arterial drip chamber120to provide a bubble-free infusion of the blood, received from the subject via first pump108, into plasma separator106. Further, EBDLR system100may include an arterial pressure transducer122to monitor a pressure of the blood entering arterial drip chamber120and provide feedback to the processor to regulate a flow rate of the blood based on the pressure of the blood entering arterial drip chamber120.

In some other examples, EBDLR system100may include a heparin pump144to inject heparin (e.g., anticoagulant) in a blood line of plasma separator106based on an activated clotting time (ACT). The heparin may prevent clogging in the plasma separator due to blood clotting. Further, EBDLR system100may include a first reservoir110to collect the plasma component from plasma separator106.

Furthermore, EBDLR system100may include a second pump112to move the plasma component (e.g., heparinised plasma component) from first reservoir110to multi-layered bio purifier102at a predefined flow rate (e.g., a predefined constant rate). Further, a threshold of 100 mmHg Transmembrane potential (TMP) is maintained. An alarm is activated when the TMP exceeds the set threshold. EBDLR system100intelligently adjusts the blood/plasma flow rate to maintain the TMP in the safe limit. Plasma is collected continuously in first reservoir110and is monitored for haemolysis using an ultra-sensitive optical blood detector. If haemolysis occurs, the cycle is immediately terminated, and an alarm is raised. Clear plasma is pumped into multi-layered bio purifier102through a sophisticated manifold (e.g., first manifold assembly146and second manifold assembly148). The manifold may uniformly split/distribute the plasma flow into the multi-layered bio purifier102.

In some examples, EBDLR system100may include at least one pressure sensor (e.g., pressure transducers132,134, and136) to monitor a flow rate of the blood and/or the plasma component and provide a feedback to a processor to regulate a flow rate in a range of 0 to 100 ml/min and 0 to 300 ml/min of plasma and blood, respectively. For example, EBDLR system100may include a first pressure transducer132to measure a pressure between second pump112and multi-layered bio purifier102. Multi-layered bio purifier102may distribute the plasma component into the first channel (e.g., first channel158inFIG.1B) and the second channel (e.g., second channel164ofFIG.1B) of each layer to purify the plasma component.

Further, EBDLR system100may include a second reservoir114to collect the purified plasma component. Detoxified/purified plasma exits from multi-layered bio purifier102via second reservoir114. In some examples, EBDLR system100may include a sampling port149to enable a user to collect the purified plasma component for testing purposes without disrupting the flow. Furthermore, EBDLR system100may include a third pump116to infuse the purified plasma component from second reservoir114into the subject. Also, EBDLR system100may include a second pressure transducer134to measure a pressure between multi-layered bio purifier102and third pump116. In some examples, EBDLR system100may include a dual filtration unit118having a 2- and 0.2-micron membrane to receive the purified plasma from second reservoir114via third pump116and filter the purified plasma to remove residual or dislodged cells from the purified plasma. In this example, the filtered plasma may be infused into the subject.

For example, the critical aspect is to infuse the purest form of the plasma component into the patient which is devoid of any residual/dislodged cells from multi-layered bio purifier102and/or bacterial contaminants. To achieve this aspect, the plasma component passes through dual filtration unit118which has a 2- and 0.2-micron membrane for filtration. The extra-pure plasma (i.e., the filtered plasma) is then infused into the subject through a bubble detector128to ensure no air bubble is injected into the patient. This cycle continues for a set volume of blood purification and detoxification. In some examples, EBDLR system100can be used continuously for long hours (e.g., 24 hours) without the need to change any of the components.

In some examples, EBDLR system100may include a venous drip chamber124to provide a bubble-free infusion of a mixture of the purified plasma component and the plasma-separated blood, from which the plasma component has been separated by plasma separator106, into the subject. Further, EBDLR system100may include a venous pressure transducer126to monitor a pressure of the mixture entering venous drip chamber124and also the subject's blood pressure and provide feedback to the processor to regulate a flow rate of the mixture based on the pressure of the mixture entering venous drip chamber124. Arterial pressure transducer122and venous pressure transducer126may provide feedback to EBDLR system100to regulate the flow rate of the blood and/or plasma component to minimally affect the blood pressure of the subject. In other examples, EBDLR system100may include a third pressure transducer136to measure a pressure between plasma separator106and venous drip chamber124that collects the mixture of the purified plasma component and plasma-separated blood from plasma separator106.

In some other examples, EBDLR system100may include an air bubble detector128to receive the mixture of the purified plasma component and the plasma-separated blood via venous drip chamber124and infuse the filtered blood and blood mixture into the subject without any air bubble. Also, EBDLR system100may include an optical blood detector130to monitor the mixture of the purified plasma component and the plasma-separated blood for haemolysis. In response to detecting the haemolysis, a process of detoxifying or purifying the plasma component may be terminated by disconnecting bioengineered dual-cell liver regeneration system100from the subject's blood circuit and generate an alarm. In some other examples, EBDLR system100may include a venous clamp147connected in a blood line. Once saline has substantially displaced all the blood from reservoir110, venous clamp147may be closed so that venous drip chamber124may be detached from the patient.

EBDLR system100described herein may use medically approved components and electronics. The sensing mechanism allows the detoxification process to happen without affecting the blood pressure of the patient. Further, the use of dual layer filter ensures that extra pure plasma is infused into the patient. Furthermore, TMP measurements and feedback ensure that haemolysis is completely avoided. Optical sensors and pressure transducers ensure that no air bubble is injected into the patient. Also, the tubing set, plasma, and capsule filters are disposable and can be used as one set per session.

Further, EBDLR system100may include the processor and memory coupled to the processor. The processor may receive inputs from different components and control the components to regulate a flow rate of the blood/plasma component flowing through EBDLR system100. The term “processor” may refer to, for example, a central processing unit (CPU), a semiconductor-based microprocessor, a digital signal processor (DSP) such as a digital image processing unit, or other hardware devices or processing elements suitable to retrieve and execute instructions stored in a storage medium, or suitable combinations thereof.

FIG.2Ais an exploded schematic diagram of an example multi-layered bio purifier200. Example multi-layered bio purifier200may include a plurality of layers202A-202N. An example layer is explained with respect toFIG.2B.

Referring now toFIG.2B,FIG.2Bis an exploded schematic diagram of an example layer202(e.g., one of the plurality of layers202A-202N) of multi-layered bio purifier102ofFIG.2A.FIG.2Cis an assembled schematic diagram of example layer202ofFIG.2B. For example, similarly named elements ofFIG.2Bmay be similar in structure, function, or both to elements described with respect toFIG.2A. Each layer202may include a membrane252having a first side254and a second side256opposite first side254. In an example, membrane252may include a biocompatible material. For example, the biocompatible material may include polycarbonate. Membrane252has a pore size of about 0.2-micron and a thickness of about 100-micron.

Further, each layer202may include a first channel260formed on first side254and a second channel264formed on second side256. In the examples shown inFIG.2B, each layer202may include a first substrate258(e.g., a first adhesive layer) disposed on first side254and a second substrate262(e.g., a second adhesive layer) disposed on second side256. For example, each of first substrate258and second substrate262may include one or more adhesive layers. An example adhesive layer may include a double-sided tape. In this example, first channel260may be defined in first substrate258and second channel264may be defined in second substrate262. Also, first channel260and second channel264may include a serpentine shape to increase a ratio of culture area to membrane252's area. The serpentine-shaped channels may provide a larger surface area for cells to grow. This can also allow for a higher amount or density of cells in first channel260and second channel264.

Furthermore, each layer202may include a first type of cells266formed on first side254in first channel260. Also, each layer202may include a second type of cells268formed on second side256in second channel264. For example, the plurality of layers may include a total number of first type of cells266in a range of 106-1010cells and a total amount of second type of cells268in a range of 106-1010cells. In an example, first type of cells266and second type of cells268are co-cultured. In the example shown inFIG.2B, membrane252may include a first membrane layer252A, a second membrane layer252B, and an adhesive layer252C (e.g., a double-sided tape) disposed between first membrane layer252A and second membrane layer252B. First type of cells266may be formed on first membrane layer252A and second type of cells268may be formed on second membrane layer252B. Adhesive layer252C may define a channel to facilitate coculturing of first type of cells266and second type of cells268.

For example, first type of cells266may include hepatocytes and second type of cells268may include endothelial cells. The hepatocytes and endothelial cells may be grown on either side of membrane252in a stoichiometric ratio of 2:1 (Hepatocytes: Endothelial Cells). Further, first channel260and second channel264of the plurality of layers202A-202N may support a maximum flow rate of 0.5 ml/min.

Further, each layer may further include a first base member270disposed on first substrate258(e.g., the first adhesive layer). First base member may include an inlet (e.g., inlets274A and274B) connected to a first manifold assembly (e.g., manifold assembly204as shown inFIG.2A). Further, each layer may further include a second base member272disposed on second substrate262(e.g., the second adhesive layer). Second base member272or first base member270may include an outlet (e.g., outlets276A and276B) connected to a second manifold assembly (e.g., manifold assembly206as shown inFIG.2A). For example, each of first base member270and second base member272comprises acrylic.

Referring back toFIG.2A, multi-layered bio purifier may include a first manifold assembly204to split incoming blood into first channel260(e.g., ofFIG.2B) and second channel264(e.g., ofFIG.2B) of the plurality of layers. The blood may include a plasma component of blood. In an example, first manifold assembly204may include a first manifold layer204A to split a first portion of the incoming blood into a plurality of first paths208A. For example, each first path may be connected to an input (i.e., inlet274A) of first channel260of one of the plurality of layers. Further, first manifold assembly204may include a second manifold layer204B to split a second portion of the incoming blood into a plurality of second paths208B. For example, each second path is connected to an input (e.g., inlet274B) of second channel264of one of the plurality of layers. Each of the number of first paths208A and the number of second paths208B is equal to the number of layers in multi-layered bio purifier200.

Further, multi-layered bio purifier may include a second manifold assembly206to combine processed blood from first channel260and second channel264of the plurality of layers. The processed blood may include detoxified or purified blood. In an example, second manifold assembly206may include a third manifold layer206A to receive and combine the processed blood from a plurality of third paths210A. For example, each third path may be connected to an output (e.g., outlet276A) of first channel260of one of the plurality of layers. Further, second manifold assembly206may include a fourth manifold layer206B to receive and combine the processed blood from a plurality of fourth paths210B. For example, each fourth path may be connected to an output (e.g., outlet276B) of second channel264of one of the plurality of layers. Each of the number of third paths210A and the number of fourth paths210B is equal to the number of layers in multi-layered bio purifier200.

FIG.3Ais a schematic top view of example first manifold assembly204ofFIG.2A. For example, similarly named elements ofFIG.3Amay be similar in structure, function, or both to elements described with respect toFIG.2A. As shown inFIG.3A, first manifold layer204A may include a first inlet302to receive and split the first portion of the incoming blood/plasma into plurality of first paths208A and a second inlet304to receive and split the second portion of the incoming blood into plurality of second paths208B.

FIG.3Bis a schematic bottom view of example first manifold assembly206ofFIG.2A. For example, similarly named elements ofFIG.3Bmay be similar in structure, function, or both to elements described with respect toFIG.2A. As shown inFIG.3B, third manifold layer206A may include a first outlet352to receive and combine the processed blood from a plurality of third paths210A and a second outlet354to receive and combine the processed blood from a plurality of fourth paths210B.

FIG.3Cis a schematic side view of example multi-layered bio purifier200ofFIG.2A. For example, similarly named elements ofFIG.3Cmay be similar in structure, function, or both to elements described with respect toFIG.2A. In the example shown inFIG.3C, multi-layered bio purifier200may have a dimension of about 120 mm in x-axis and y-axis and about 20 mm in z-axis.

Thus, multi-layered bio purifier200described herein may provide a systematic array of 2 cell types which function as a mini-liver. Multi-layered bio purifier200may be a multi-layered microfluidic platform. Each layer is composed of 2 channels. The channels are isolated by a customised and selectively permeable membrane (e.g., polycarbonate; 0.2-micron pore size, 100-micron thickness; coated with poly I lysine). Hepatocytes and endothelial cells are grown on either side of this membrane in a stoichiometric ratio of 1:2. This may be the optimum ratio for maximum detoxification efficacy. The cells cross-talk with each other and complement the detoxification functions. A state-of-the-art manifold assembly is integrated in multi-layered bio purifier200to split the incoming plasma flow into multiple channels. A similar manifold is present at the other end of multi-layered bio purifier200to collect the processed/detoxified plasma.

The multi-layered bio purifier200is designed and developed to grow 2 different types of mammalian cells, for instance, hepatocytes and endothelial cells. These cells when grown together, complement each other's functions and therefore the entire assembly works like a mini liver. A unique, customized membrane is used in multi-layered bio purifier200which allows the user to grow 2 different types of cells. Multi-layered bio purifier200has flow paths/channels which support a maximum flow rate of 0.5 mL/min. Multi-layered bio purifier200may support the growth and residence of up to 1010cells in either channel. The unique character of multi-layered bio purifier200is that this arrangement mimics the architecture of cellular lining and blood vessels in the human body. Multi-layered bio purifier200can be used multiple times with appropriate cleaning and sterilization. Growth conditions of the cells in multi-layered bio purifier200may include: 37-degree Celsius, 5% CO2 and 95% relative humidity. These conditions are optimal for the cells to perform their functions. Multi-layered bio purifier200design has been made to grow any kind of mammalian cell. Currently hepatocytes and endothelial cells have been grown. In the future, multi-layered bio purifier200can be used to grow different cells like B cells and the like, which can aid in developing a device for diabetes management.

FIG.4is a flow diagram illustrating an example method400for treating a subject with acute liver failure. Example method400depicted inFIG.4represents generalized illustrations, and other processes may be added, or existing processes may be removed, modified, or rearranged without departing from the scope and spirit of the present application.

At402, an extracorporeal bioengineered dual-cell liver regeneration (EBDLR) system may be included into the subject's blood circuit. The blood circuit may refer to a blood line for transporting blood between a patient and the EBDLR system. At404, blood may be passed through the EBDLR system. In an example, the blood may be continuously taken from the subject. In an example, passing the blood through the EBDLR system may include providing a bubble-free infusion of the blood taken from the subject via a first pump into the plasma separator using an arterial drip chamber. Further, a pressure of the blood entering the arterial drip chamber may be motored using an arterial pressure transducer and a feedback may be provided to a processor to regulate a flow rate of the blood based on the pressure of the blood entering the arterial drip chamber.

At406, the blood may be separated into a plasma component and remainder using a plasma separator of the EBDLR system. For example, the blood may be passed through the plasma-separation element with a flow rate of 0-300 ml/min. In an example, heparin may be injected in a blood line of the plasma separator based on an activated clotting time (ACT) using a heparin pump. The heparin may be to prevent clogging in the plasma separator due to blood clotting.

At408, the plasma component may be passed into a bio purifier having a plurality of layers. In an example, each layer may include hepatocytes on a first side of a membrane in a first channel and endothelial cells on a second side of the membrane in a second channel. For example, the plasma component may be passed through the bio purifier with a flow rate in a range of 0-100 ml/min.

In an example, passing the plasma component into the bio purifier may include:collecting the plasma component into a first reservoir,passing the plasma component from the first reservoir into the bio purifier via a second pump, andmonitoring a pressure of the plasma component passing into the bio purifier using a first pressure transducer and providing a feedback to a processor to regulate a flow rate of the plasma component.

At410, the plasma component may be split into the first channel and the second channel of the plurality of layers to purify the plasma component. In an example, splitting the plasma component into the first channel and the second channel of the plurality of layers may include:splitting, using a first manifold layer, a first portion of the incoming blood into a plurality of first paths. In an example, each first path may be connected to an input of the first channel of one of the plurality of layers.splitting, using a second manifold layer, a second portion of the incoming blood into a plurality of second paths. In an example, each second path may be connected to an input of the second channel of one of the plurality of layers.

At412, the blood may be restored by combining the purified plasma component and the remainder. After plasma has been purified, the blood is restored by combining the purified blood plasma and the remainder and the restored blood is continuously returned into the patient.

At414, the restored blood may be continuously returned into the subject. Further, the EBDLR system may be separated from the patient's blood circuit upon performing the blood purification process for a predefined number of days (e.g., 28 days). In an example, continuously returning the restored blood into the subject may include providing a bubble-free infusion of the restored blood into the subject using a venous drip chamber and an air bubble detector. Further, a pressure of the restored blood entering the venous drip chamber may be monitored by a venous pressure transducer a feedback may be provided to a processor to regulate a flow rate of the restored blood.

In an example, continuously returning the restored blood into the subject may include monitoring, using an optical blood detector, the restored blood for haemolysis. In response to detecting the haemolysis, the EBDLR system may be terminated from the subject's blood circuit and an alarm may be generated.

In an example, method400may include passing the purified plasma component through a dual filtration unit to filter the purified plasma component. For example, the dual filtration unit may include a membrane to devoid any residual/dislodged cells from the bio purifier and/or bacterial contaminants from the purified plasma component. The blood may be restored by combining the filtered plasma component and the remainder. The purified plasma component may be passed through the dual filtration unit with a flow rate in a range of 0-300 ml/min.

In an example, passing the purified plasma component through the dual filtration unit may include collecting the purified plasma component from the bio purifier into a second reservoir. Further, the purified plasma component may be passed from the second reservoir into the dual filtration unit via a third pump. Furthermore, a pressure of the purified plasma component passing into the dual filtration unit may be monitored using a second pressure transducer and a feedback may be provided to a processor to regulate a flow rate of the purified plasma component.

Thus, the EBDLR system described herein functions like a liver. Detoxification capabilities of the EBDLR system can be used to treat a subject with an acute liver failure. The EBDLR system functions without affecting the haemodynamic equilibrium of the patient. Haemolysis-free plasma is processed/detoxified through the bio purifier. The multi-layered bio purifier uses bioengineered components (e.g., cells) to detoxify/purify diseased plasma. Uniform flow distribution is achieved via integrated manifold assemblies in the bio purifier assembly, which is developed by employing principles of Murray's laws and biomimetics. The biocompatible membrane for the cell culture is customized as per the need and is made of polycarbonate material with a pore size of at least 0.4 microns. Effective detoxification happens at a flow rate ranging from 0.2 to 0.4 ml/min. These flow rates correspond to a shear stress range of 200 to 500 dyne/cm2. The EBDLR system draws blood from the subject without affecting the blood pressure and does not introduce turbulence in the native blood flow. Various pressure transducers enable the user to identify various flow parameters and optimize the operations. Once primed, the EBDLR system runs continuously in an automated manner. The EBDLR system is easy to setup, can run continuously for 24 hours and has a capability to purify the complete blood volume from a patient's body. The EBDLR system has proved to work with 100% efficacy in the porcine model of acute liver failure.

Results From Large-animal Trial Using the EBDLR System

Exhaustive animal trials were performed using the EBDLR system to evaluate the efficacy of the EBDLR system. Briefly, the acute liver failure was induced in Yorkshire pigs using D-galactosamine. 24 hours post induction, the animals were connected to the EBDLR system for 4 hours. Blood was detoxified using the EBDLR system. The device connection procedure was done in the animals without anaesthesia. In this case, haemodynamic stability was maintained without anesthesia. This process was repeated after 48 hours.

The blood glucose monitoring and maintenance is of critical importance for an individual undergoing treatment with the EBDLR system. With the examples described herein, a no-infection condition may be achieved during the entire course of experiment which lasted for 28 days. During the course of the study, the bio purifier and plasma filter were replaced at regular intervals (e.g., 24 hours) whereas the rest of the EBDLR system was thoroughly decontaminated and reused for the subsequent cycles. A detailed record of clinical investigations was maintained. This included blood glucose, liver function parameters, kidney function tests and infection checks.

The overall outcome of the experiment was that the animals connected/treated with the EBDLR system post liver injury, recovered fully in 28 days whereas, the control/untreated animals succumbed to acute liver failure in 36-48 hours.

The above-described examples are for the purpose of illustration. Although the above examples have been described in conjunction with example implementations thereof, numerous modifications may be possible without materially departing from the teachings of the subject matter described herein. Other substitutions, modifications, and changes may be made without departing from the spirit of the subject matter. Also, the features disclosed in this specification (including any accompanying claims, abstract, and drawings), and any method or process so disclosed, may be combined in any combination, except combinations where some of such features are mutually exclusive.

The terms “include,” “have,” and variations thereof, as used herein, have the same meaning as the term “comprise” or appropriate variation thereof. Furthermore, the term “based on”, as used herein, means “based at least in part on.” Thus, a feature that is described as based on some stimulus can be based on the stimulus or a combination of stimuli including the stimulus. In addition, the terms “first” and “second” are used to identify individual elements and may not meant to designate an order or number of those elements.