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With the growing area of regenerative medicine, it’s no surprise that there are dozens of systems available to process Platelet Rich Plasma (PRP). While the use of each system will result in a plasma product, the processing techniques, equipment and final sample differ quite drastically. In this installment of our Myth Buster series, we will take a closer look at the differences that one can expect to encounter when looking at PRP processing systems and how each of these components effects the PRP product.

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Processing Techniques

There are three main methods by which Platelet Rich Plasma can be processed. These methods are:

1. Centrifugation – This is the most common technique available, which involves using a centrifuge to separate the cell layers in whole blood or to pass the blood components through a gel separator (which is in the device prior to insertion of blood) to isolate the platelets from other cell types.

a. Blood Layer SeparationSpinning without gel separator – When blood is spun at a high rate of speed for a period of time, the cellular components will separate according to their density within the container/tube. These cell layers can be visualized upon removal of the device as seen in the below picture. The three distinct layers of blood include the plasma layer, the platelet buffy coat and the red blood cell layer. The plasma and buffy coat can be collected and injected or the suspension can be placed in a second container for a second spin. The second spin in the process enables further concentration of the platelets, which as we discussed in our previous posts, is important for delivering a therapeutic concentration of platelets (between 3-7-fold concentration compared to circulating blood). A sample that is obtained by a single spin process will have a lower concentration of platelets, thus possibly resulting in a sub-optimal therapy.

b. Spinning with gel separator – Gel separators provide a physical gradient for blood to pass through resulting with the final product at the top of the tubes. Gel separators are typically used for isolating either plasma or serum, but have been developed to allow smaller cell types to remain in the final product. Unfortunately, the majority of products available that use gel separation produce a low concentration of platelets due to the small volume of blood being used (typically 8- 10 mL) as well as the poor capture of platelets in the final product.

2. Flow Cytometry - This method involves a highly specialized piece of equipment that utilizes the absorption of light by a cell and further separates the cells according to the refracting light. This process relies on blood flowing in a single layer suspension through a complex network of tubes, which can be further complicated if the sample begins to clot during the process. Flow cytometry is typically automated, which provides great convenience to the user, but the equipment may require additional set up (prior to processing) and upkeep to ensure proper calibration for processing samples.

3. Gravity and Reverse Osmosis – This method involves utilizing a specially designed blood containment system with a built-in filter. The filter’s purpose is to collect the platelets while allowing the other cells to pass through. Although this method is attractive from a space and equipment perspective, performance of these systems has shown to result in an increase of neutrophils which can be detrimental when injected into a joint.

 

Volume of Blood

Blood compositionOne of the biggest differences between PRP processing systems is the volume of blood that is required to obtain the PRP. Some systems require as low as 8 mL of whole blood, while other systems, such as the CRT system, require between 25-50 mL. What does this variation mean to you? If we look at typical blood composition, as depicted in the picture below, we will see that platelets make up less than 1% of the circulating blood. In order to have a therapeutic dose of platelets in the volume which is necessary for administration, a higher volume of blood is required for processing. For example, when using the CRT system, it is recommended to collect 50 mL of whole blood to produce 4-5 mL of PRP (with concentration of platelets between 3-7 fold). For systems that require the smaller whole blood volume with the same end product volume, the concentration of platelets will be below the recommended therapeutic threshold.

 

Sterility/ Exposure to Environment

When considering any product that will be administered intra-articularly or into areas of poor blood flow, it is imperative to ensure that the processing is done with aseptic technique. It is also important that the exposure of the sample to environmental factors is minimal to reduce the likelihood of contamination. The majority of systems available are considered to be closed systems in which the sample being processed has little to no contact with the outside environment. There are some processing systems available which utilize open tubes or require passing the blood through a needle into the containment device, which may introduce additional contaminants into the sample.

Time to Process- From Collection to Administration

Depending on the technique of isolating the platelets (centrifuge vs. flow cytometry vs. gravity filter), processing times vary greatly from just under 10 minutes to as much as 45 minutes! Some processing systems require activation of the platelets, which can take up to 45 minutes. Activating platelets, however, is not a necessary step, since the platelets will activate and release their stored growth factors once they are exposed to collagen in the joint/tissue. The Companion Regenerative Therapies System takes less than 15 minutes from blood collection to administration, making it a fast, in-house therapy that can be easily scheduled even in a busy hospital.

 

To learn more about Platelet Rich Plasma and how it works as a therapy, watch this short animation:

Stay tuned for our next blog post where we “bust” another regenerative medicine myth!

 

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In the veterinary hospital, a major component revolving around the integration of a new therapy is time. How long is the treatment going to take? How many personnel will need to be scheduled to administer the therapy? How long does the patient need to be under sedation (if needed)? How long does the patient need to be in recovery?

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The old saying “time is money” certainly has its place in the veterinary hospital. If a procedure requires several technicians and takes numerous hours to perform for treating one patient, that may not be an efficient use of the hospital’s or technician’s time. In the early years of regenerative medicine, many of the therapies required numerous anesthetic episodes and several hours of both the technician’s and doctor’s time. The procedures required a dog to be anesthetized to collect the sample and then return in a separate hospital visit to be administered the therapy. The collection of the tissue and administration required the doctor to be scheduled for two surgical procedures as well as the attending technician(s). In total, the time to perform this therapy could easily approach 2 hours of doctor and technician time combined, which would then necessitate a higher fee for the single therapy to cover the overhead costs. For many customers, the combination of the numerous office visits and cost of the procedures would be prohibitive, making this therapy less appealing.

Today, these therapies have become extremely efficient only requiring a single office visit with processing and administration taking less than 30 minutes. For Platelet Rich Plasma (PRP) the process takes even less time (typically 15 minutes from start to finish) with most of the procedure being performed by the attending technician(s). The steps for processing PRP are quite simple, making it a therapy that can be easily cross-trained for the supporting staff. The doctor’s time is only required for the administration of the therapy, which takes less than 2-3 minutes (depending on the number of areas being injected). With this minimal time requirement, providing regenerative therapies can easily be scheduled in-between other procedures, exams or surgeries.

To hear a first-hand account of how the Companion Regenerative Therapies System benefited a general practice and their schedule:

Stay tuned for our next blog post where we “bust” another regenerative medicine myth!

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Spinal cord injuries can have a devastating impact on a patient’s life. For the canine patient, spinal cord injuries are surprisingly common, accounting for approximately 2% of all cases that are presented to the veterinarian*. The most common forms of injuries arise from acute disc herniation, spinal trauma, fibrocartilaginous embolic myelopathy and chronic diseases including degenerative myelopathy and spinal stenosis. While the initial injury to the spine is a major concern, it’s the secondary effects of that trauma that have long lasting effects including impaired blood flow which can lead to “stroke-like” events.

Spinal tissue has poor regenerative properties, which can provide a challenge to the attending veterinarian when choosing a treatment program for their patient. Depending on the severity of the injury, some patients can make a significant recovery with a comprehensive rehabilitation program. Some patients, however, may remain unresponsive to physical rehabilitation and require further intervention. Below is an overview to a case study originally reported in Veterinary Practice News  (February 2016) by David R. Mason, B. Vet. Med, DACVS, DECVS, of Las Vegas Veterinary Specialty Center. This case study focuses on a young, 5-year-old Labrador Retriever named Duncan with Fibrocartilaginous Embolic Myelopathy (FCE).

FCE is a condition in which a fragment of disc material is somehow forced into the spinal vasculature. This migration of disc material causes a “stroke-like” event, cutting off blood flow to the surrounding spinal tissue. This event can cause partial or complete paralysis to the patient. FCE is most commonly reported in large and giant breeds, but is also prevalent in sheep dogs and miniature schnauzers. The most common treatment protocol for patients with FCE is based around aggressive supportive care and physical therapy.

Duncan’s owners reported that he was in the back yard when he vocalized and then was found unable to move his back limbs. After undergoing a thorough evaluation at Las Vegas Veterinary Specialty Center (LVVSC), it was determined that Duncan was suffering from FCE. Subsequently, Duncan was enrolled in a 16-month rehabilitation program, but his condition did not improve and he remained non-ambulatory in his back limbs. Knowing that additional treatment options were limited for Duncan, Dr. Mason elected to treat Duncan with a combination of Platelet Rich Plasma (PRP) and Bone Marrow Aspirate Concentrate (BMAC). Duncan was treated with an epidural injection of PRP/BMAC solution which was produced using the Companion Regenerative Therapies System. After injection, Duncan recovered from anesthesia without incident and returned to his dedicated rehabilitation program.

By day 14, Duncan’s owners reported an increase in energy and after continuing his rehabilitation, Duncan is now able to take several steps without the assistance of a sling or cart. Although Duncan’s condition may not return fully to pre-injury form, this case provides a positive outlook to treating spinal conditions with regenerative medicine.

To read more about Duncan’s story, click here.

To learn about the Companion Regenerative Therapies System and upcoming educational opportunities, please click here.

* https://cvm.ncsu.edu/research/labs/clinical-sciences/canine-spinal-cord-injury/

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