Hello, thank you to those that are joining us live and for those that will access this recording. Today's webinar is titled Regenerative Medicine and Veterinary Health, Stem Cell and PRP Therapy. Quick housekeeping to mention.
If anyone has any questions for our speaker, please put those into the Q&A box, and we will cover those at the conclusion of the presentation. For comments or other queries, you can use the chat box and we'll address that live. Just a quick bio on our speaker today who is the site leader for Arent Animal Health located outside of Lexington, Kentucky.
Doctor Adrian Wright has a profound passion for stem cells and regenerative therapies. Doctor Wright brings both expertise and heart to Arent's mission. A proud University of Kentucky graduate with a bachelor's degree in chemistry.
Adrian also holds an MBA in data analytics and a PhD in anatomy and physiology from Kansas State University College of Veterinary Medicine. Both of which she completed concurrently. Her journey spans lab research to leading analytical teams, budgeting rigorous science with strategic business insight.
Turn things over to you, Doctor Wright. Thank you for the introduction, Josh. I started my career at Ardent and I came back to Ardent a little over a year ago because I believe in this company, and I believe in regenerative medicine.
I have close to 15 years of experience in regenerative medicine, and I feel that it is a valuable tool for veterinarians to help treat patients that this is maybe their last resort. Or even just to give your patients other options. Even further, adding regenerative medicine to your treatment protocols allows you to diversify your practise from other practises in your area and boost revenue for the practise while staying on the innovative side of care.
Today, we will dive into the regenerative medicine areas that Arent can help provide to your practise. The first idea of regenerative medicine appeared in publication roughly 30 years ago, and it was predicted as a new branch of medicine will develop that attempts to change the course of chronic disease and in many instances will regenerate tired and failing organ systems. This prediction was made not by a clinician, but by a hospital administrator back in 1992.
He believed a new branch of medicine might come about and change the way chronic diseases are looked at. But the idea of modern regenerative medicine as we know it today, didn't appear in publications until almost a decade later. Since then, regenerative medicine has become a key component of modern medicine.
Rapid advancements in cell therapy, tissue engineering, gene therapy, and other biologics have completely changed the way we treat conditions that were once thought to be untreatable. Starting with platelet-rich plasma or PRP, this product is made from the patient's whole blood. The basis of PRP is really pretty simple.
You're concentrating the platelets above the baseline whole blood value. From those concentrated platelets, growth factors are released from the alpha granules within the platelets. These growth factors then are involved in signalling the cells in the body to migrate to areas of injury to promote tissue healing and reduce inflammation.
Platelets are known to be involved in cell migration, cell differentiation, angiogenesis, cell to cell adhesion, and cell signalling. PRP is known for its widespread use, but it is most prevalent in orthopaedics, dermatology, dentistry, and aesthetics. The first clinical reports of use in PRP date back to the 1980s, and since then use has exploded.
Once the platelets are concentrated, the platelets must be activated or laced to release the growth factors from the alpha granules. You don't want to rely on the body to completely activate or lice those platelets once they're injected. This can be done either chemically or mechanically.
Argen uses a chemical activation and it can be the most time efficient mechanism. It's done in a single step. Mechanical activation typically involves multiple rounds of freezepa cycles to lice the platelets through an ice crystal formation.
This can be quite laborious and is spread over multiple hours, so it's really not ideal for in clinic protocols. The growth factors that are released include, but are not limited to VeF, FGF, IGF, TGF Beta, PDGF probably the most prominent, and EGF. But this list goes on and on, and we still do not know every single component of PRP at this time.
Not all PRP preparation systems are the same. Dr. Sam Franklin at the University of Georgia did some canine PRP characterization work on PRP kits.
He tested 5 commercially available kits, and ardent kit was one of the 5 tested. We are kit number 2, just for your knowledge, but all kits are identified in the paper, and I'm more than happy to send this paper to anyone that's interested. He used 15 healthy donor dogs with all of the kits, and there was variability in all parameters tested.
So that whole blood starting volume ranged from 9 mils up to 55 mLs. This is important to note because it can be difficult to get larger volumes from smaller dogs. The PRP volume that was obtained at the end ranged from 2.9 mils to 6.2 mils.
And you might want to think that the larger volume is best, but it really depends. You still want therapeutic concentration. So a rule of thumb is 3 to 5 times the baseline.
So knowing your concentration is important, but you do want to make sure that you have a sufficient volume at the end for all your injections, so that you don't end up needing two kits. Speaking of platelet concentration, which this is the therapeutic part of the PRP. This actually only occurred in 4 of the 5 kits that he tested with kit number 1 actually decreasing the concentration from baseline, which does not meet the definition of a PRP.
The other four kits ranged from 1.8 to 5.2 times the baseline.
Leukocyte concentration was also highly variable, so 2 systems decreased the leukocyte concentration, while the other three concentrated the leukocytes. And I know that from this data, the idea of a PRP kit can be quite overwhelming. So it really is important as a clinician to understand the goals of what you're treating and what constraints you may have, such as a limited starting blood volume or maybe a high volume need at the end for the injections when you're choosing a system.
So now to the great debate or sort of what's trending in PRP right now. Leukocyte poor versus leukocyte rich PRP. If you remember from the previous Franklin paper, two systems decreased the leukocyte, which gives what we call a leukocyte-poor product, while the other three systems concentrated the leukocytes, which will provide a leukocyte-rich product.
So, which one's better and what's the difference? Well, the answer is it depends. Researchers and clinicians seem to feel strongly about this with little evidence to really say which one is definitively better, or even any evidence that has been replicated.
So you have the leukocyte-rich PRP. So the contents would be the leukocytes, the platelets, and the plasma. Some do consider it to be enhanced.
Others argue that the white blood cells, which are known to be inflammatory, might flare inflammations in an uncontrolled manner. Some veterinarians hypothesise that this is more ideal for chronic conditions where inflammation may be lower. So by using a more pro-inflammatory product, you are actually turning a chronic condition to an acute setting.
We do see this used a lot in tendinopathies. Then you also have leukocyte poor PRP. So the white blood cells are reduced, so your main components are really just platelets and plasma.
It does have the ability to deliver a more controlled inflammatory response due to the anti-inflammatory nature of the product. It is ideal for acute conditions where inflammation may already be super elevated and you really don't want to elevate that inflammation any further. Some studies have shown superiority to this version, while others have shown no statistical difference between the leukocyte poor when you compare it to leukocyte rich.
I do want to point out that Ardent's PRP kit is written as a leukocyte pore formulation, but it can be made into a leukocyte rich with some really easy protocol modifications that our team can walk you through. Ardents in clinic and lab service PRP option is super simple to run. It takes about 1 hour total of tech time.
It is an affordable option where the kit costs about $130 and the lab service runs $300 which includes shipping and processing by the ardent team. Briefly, the blood is collected and centrifuged. You collect the buffy coat and centrifuge again to concentrate your final volume.
The platelets are activated to release those growth factors that we want. Again, we use a chemical activation method. The concentrated platelet plasma will form a fibrine clot.
This clot can be used topically for burns and other wounds. Otherwise, you wait for the liquid to retract from the clot and it's ready to inject. On the right, you can see all of the uses for PRP, but we are honestly discovering more every day.
So we have ocular. I'm currently using it for KCS with my own dog. We have oral issues like stomatitis, which you'll see in a few minutes.
We have arthritis, surgical repair. I do want to tease a paper on PRP with surgical repair that's gonna be coming out in the next couple of months from Doctor Darryl Milli at the University of Tennessee. So be on the lookout for that.
We also have burns and wounds, tend tendon injuries, cruciate tears, just to name a few. Arent sells an equipment bundle that comes with a stock of consumable kits to generate a return on your investment. Or some practises may have an existing swinging bucket rotor centrifuge that will work and they're able to simply purchase the kits.
Josh, who you heard from earlier, can help determine if your current machine will work. We have selected a few of our favourite PRP case studies. This one is a feline stomatitis case coming from Dr.
McMillan at Braden Run Animal Hospital. So the before photos of Maverick are on the top and the bottom is Maverick's mouth 7 days post injection. Doctor McMillan essentially used a tattoo method where she injected the oral lesions many times with small PRP volumes using a 30 gauge needle.
They did end up doing a second treatment for Maverick, but it wasn't until 5 months later. No dental extractions were needed. Some may need injections sooner.
This really is gonna depend on the animal. But the inflammation and lesions have substantially decreased. I really wouldn't even think this is the same cat.
But as Dr. McMillan points out, PRP is a great tool for owners who cannot afford a full mouth extraction, stem cell, or other treatments for feline dermatitis. So then we have Rudy.
Rudy received a total of 3 PRP injections, about 30 days apart because he is an older patient with advanced arthritis. A normal arthritic patient may get 1 injection in around 3 to 6 months of improvement. But again, this is gonna be dependent upon the animal.
But I really want you to take advantage of that frozen PRP. PRP can actually withstand a single freeze-thaw cycle and be stored in a standard freezer at your hospital for 90 days. Next we have Chloe.
Chloe was an IVDD dog treated at Good Shepherd Pet Hospital. Doctor Deeb had tried prednisone, gabapentin, and carprofen prior to reaching for PRP. He did the first injection in October of 2021 and the second in July of 2023.
His protocol was a half mil intrathecal with 0.3 mLs in six injection sites, they did a zigzag pattern around the spine. We have a feline trauma case.
This one used PRP with a cold laser. This pregnant cat came in after being hit by a car with a fractured pelvis, unable to walk or go to the bathroom, scheduled for euthanasia by the local Humane Society. They were referred to Doctor Rashkovic at Golden Bone Wellness centre for evaluation.
She treated with PRP and cold laser therapy. She injected that PRP around the fracture sites, and within 24 hours, this cat was able to walk and go to the bathroom on her own. One week later, she naturally delivered 1 kitten and the second was delivered by C-section.
We had a full recovery for the mom. Both kittens were adopted, and this cat currently still lives with Doctor Rashkovic. My favourite P PRP case is my own dog.
This is Blanche. I adopted Blanche earlier this year from a rescue in Tennessee. She had been born in Alabama.
She had had her initial care there. She'd had her cherry eye surgery. From there, she was surrendered.
She bounced around to a couple of other states, went into some foster homes before I rescued her. So her care had just been very sporadic and tough to follow. But due to that cherry eye surgery, she was actually diagnosed with KCS in her right eye back in April.
My veterinarian immediately prescribed steroid eye drops, but it's not really something that I wanted to continue for the rest of her life, so I encouraged him to try PRP. Within a week, Blanche had increased tear production in her right eye and was comparable to slightly lower than the left. I continue to do 1 to 2 drops in the affected eye in the morning and in the afternoon for a total of 4 weeks.
I definitely was not a perfect pet owner here, which is what makes this case so nice. This is the adherence to the protocol that you'll probably see with a typical owner. But Blanche's eyes were immediately less irritated.
She loved, she rubbed them less, and the eye secretions drastically dropped off. When I first started giving her the treatments, she resisted. She wasn't really excited about it, but in the end, it's like she knew that it was gonna make her eyes feel better and she would easily let me do it.
But following the treatment at the end of the 28 days, we took a final photo in a tear production reading, and since then, we've just continued to see how she's maintaining. Her sclera is less red and irritated. Her tear values have increased, eye secretions have not returned.
She's maintained this state for a couple of weeks following the end of treatment, and she is on zero medications for the KCS. The corneal edoema that you see in the first picture is almost completely gone. And then since then, my veterinarian has actually actually treated a couple of more KCS cases with PRP just due to Blanche's results.
We were actually just sent this case from Dr. Weiss at Doylestown Vet Hospital. So Della came into the clinic on April 20th with an open wound following a dehance of surgical incision to remove a cancerous mass.
The clinic used a single treatment of the gel clot from the PRP kit. It can also be called PRF. They were not able to bandage due to it being in an awkward location where the bandage wouldn't stay.
This is the photo of her on the bottom left of your screen on April 20th, that's the day zero. You can see the wound on the top right, taken just 9 days later, on April 29th. You can see the wound has made tremendous progress in the healing journey and is starting to close.
The last photo we have is on the bottom right, it's on May 19th, almost one month later. The wound is almost completely closed. And again, this was from a single treatment of PRP with the kit.
If you're interested in diving more into PRP's role in wound healing, I do have a podcast episode where I review a case study of wound healing using PRP and the antimicrobial effects of PRP. The podcast is called Sit Stay, Learn, and it's available wherever you listen to podcasts, or we can send you a link. PRP works great for ocular issues.
This is a case of a corneal ulcer treated with PRP by Doctor McKenzie at Pet Life Vet Hospital in Trinidad and Tobago. You see before on the left, for this case, Dr. McKenzie did 2 drops of PRP 5 times a day.
30 minutes after the PRP was administered, they did a topical antibiotic drop and they repeated this procedure daily for 7 days, and then they were scheduled for a recheck. The owner actually bailed on the recheck stating that the eye was great, it's returned to normal, and they didn't need anything else. They did finally get the owners to come in, so that's when you see the after photo on the right, that's actually day 18.
The eye looks amazing and no further treatment was needed. PRP can be used as a standalone treatment, or it is used in conjunction with stem cell therapy. So when someone thinks of regenerative medicine, stem cells are one of, if not the first thing that comes to mind.
And when they say stem cell, they are typically referring to meinomal stromal cells, or MSCs. We'll go into those types in just a minute. But the concept of stem cells actually originated at the end of the 19th century, but they were first described by Friedenstein and his group in 1968 as a population of fibroblast-like hematopoietic support cells in the bone marrow.
MSCs as we know them today, weren't described until 1991 by Arnold Kaplan, and from there they've taken off. The discovery of stem cells in the late 1960s marked a pivotal moment in science that confirmed the idea from the 1800s that the body does have the ability to regenerate and repair itself. This work by Friedenstein and his colleagues laid the foundation for modern regenerative medicine and has transformed the approach that we take for treating injury and disease across both human and veterinary fields.
So, what actually is a stem cell? Well, I myself am guilty of using this generic term, although it actually encompasses many cell types. Basically, a stem cell must be undifferentiated, and it must be capable of two very important properties.
So one is self-renewal. So it has to be able to make more stem cells. They will also make more progenitor cells, but that being able to make more stem cells is the most important.
Also, there's differentiation. So there are different levels of differentiation required for different potency of stem cells. Stem cells vary based on their potency, and when we say potency, we're talking about how strong they are or how stimmy the cell is.
So you start at the very top with totipotent cells. They can divide and produce the entire organism, including embryonic and the extra embryonic tissues. So a zygote is totipotent.
Then you have the pluripotent cells. These are your embryonic stem cells, they can produce all of the germ layers. So the ectoderm, the mesoderm, and the endoderm.
These cell types come with a lot of challenges, both scientifically and ethically. They are really quite challenging to work with, but they have been maintained for decades as the gold standard. You can also have IPSCs, which is a newer discovery.
Those are essentially multipotent cells that are then reprogrammed to be pluripotent again. But again, they come with their own challenges, they're difficult to work with. We then have multipotent cells, and this is what most people are talking about when they say stem cells.
They are adult stem cells, and they can differentiate into multiple cell types. These are what we simply call MSCs. We also have unipotent cells, and these are already committed cells that can only differentiate into one cell type.
MSCs are plastic adherent multipotent progenitor cells that are capable of differentiating into cells of the mesodermal lineage. To meet the International Society of Cell Therapy definition of minimally what an MSC is, they must be able to differentiate into bone, fat, and cartilage. That's not to say that they can't differentiate into more, but a lot of the studies out there are just gonna demonstrate these three, so they can say they have an MSC.
They also have to have a distinct surfer distinct surface marker panel of both negative and positive markers that must apply to greater than or equal to 95% of the population for them to be called an MSC. The sources include the bone marrow, which is where they were discovered by Friedenstein, so that's why you see those as maybe the more popular choice. But there's also the adipose tissue, the umbilical cord, dental pulp, placenta, muscle, and many more.
The therapeutic potential of MSCs is quite vast, and honestly, we still don't know everything about these cells. We don't know their full potential and we don't really know their full limitations. They are most often recognised for their migration, homing, and tissue repair capabilities, but they're also immunomodulatory, and they also have low immunogenicity.
We used to believe that they were immune privileged, but we now confidently know that they are not. They are surveilled by the innate immune system. Ardent animal health uses the adipose tissue from cats, dogs, horses, and many exotics.
But why do we use the adipose tissue? Well, it's a rich source of MSCs. Adipose tissue, gramme per gramme contains roughly 100 times more MSCs compared to that same amount of bone marrow.
It is an accessible source of MSCs, so it can be collected simply, and it's relatively painless. Most animals also have an abundance of adipose tissue, and you can very easily get more adipose tissue than you can get bone marrow. And the MSCs from these two sources have been shown in multiple studies to be similarly effective.
I think the only reason that bone marrow was still holding out is because it was the original source where they were discovered. Arent animal health isolates their product from the adipose tissue, and although it is rich in MSCs, the primary isolate that is obtained is not pure MSCs. It's actually known as a stromal vascular fraction or an SVF.
But the SVF is a heterogeneous mixture of cells and growth factors that are extracted from the adipose tissue, usually following an enzymatic digestion, but it can also be mechanical. It is a very rich source of MSCs, but there's also endothelial precursor cells, Tregs, macrophages, smooth muscle cells, in addition to many other growth factors, cells, and proteins. But this population plays a role in signalling, regeneration and repair.
SVF is currently used for cosmetics, aesthetics, immune disorders, orthopaedics, cardiac, neurological, wound healing, and even diabetes. The benefits of SVF are that it's quick, so the cells can be isolated and injected back into that patient in the same day. There's no time in culture, so sometimes culturing can take weeks.
This is gonna reduce cost and the potential for contamination. There's also a reduced risk of allergens to ingredients that are involved in the cell culture, such as FBS. And this heterogeneous pool of cells, although it's not fully characterised, does work together to create an incredible clinical benefit.
Ardent has been treating dogs with OA and other conditions with their SVF for over 15 years. In 2016, we published a safety and efficacy study that was completed on client-owned dogs at Kansas State University. 22 dogs were treated, 12 placebo, and 10 stem cell.
This was placebo controlled and double blinded. The dogs must have had OA of the hip joint in an otherwise healthy condition. They were evaluated with a physical exam, lameness score, a pain index survey, and a force plate analysis on a pressure sensing walkway.
They were treated with a single intra-articular injection and an IV dose. They were evaluated at day 0, and then weeks 48, 12, and 24. Dogs that were treated with the SVF had improvements in pain and were able to put more weight on the affected limb as seen by the peak vertical force.
No adverse events were reported, and at the end of this study, fewer dogs in the treatment group had lameness compared to those receiving the placebo. When you look at osteoarthritis treatment in canines with SVF or MSCs, multiple studies have shown improvement in dogs, even long-term improvement. In a study by Macky, using an intra-articular injection of adipose derived MSCs, pain and lameness was reduced for dogs suffering from hip osteoarthritis.
They treated 20 dogs, they used a saline placebo, and they varied the dose of the cultured MSCs. Pain reduction was evident in all of the MSC treated groups, but surprisingly, they had the greatest reduction with the lowest dose of MSCs, which was 5 million cells. When breaking down just MSC treated to placebo treated, roughly 60% of dogs had a pain reduction compared to 20 of those treated with placebo.
And when you look at the lameness scores, in the placebo group, the average lameness score was a 0.5, compared to around a 2 for MSC treated dogs, showing that MSC dogs did have a significant improvement in lameness compared to those dogs treated with saline. Again, no serious adverse events reported.
In another study from Bruins, this tested a single injection of SVF in canines for elbow osteoarthritis. They had 23 dogs. All dogs were unresponsive to conventional therapy.
In 19 of the 23, joints no progression of OA was noted. Peak vertical force improved significantly at 3 months. At 6 months, 26% of dogs had improvement in pain, and 67% at 12 months.
Again, no adverse events reported. But what can MSCs be used for? Again, we really don't know the full capability or the limitations of these cells, but new papers are coming out daily.
So MSCs and particularly, I'm gonna look at adipose derived, have been linked to studies showing use in hip dysplasia, atopic dermatitis, diabetes, IBD, IVDD, and ligament tears. And clinically, what we are seeing is astounding. I'll let the case studies speak for themselves.
We're gonna present some basics for the cases like we did with PRP, but if you see any particular case that interests you, please reach out. We can gather more details for you. We can get you in contact with our technical services veterinarian, Doctor Larry Snyder, who can give you some case guidance.
Maggie is a 9 year old mixed breed with bilateral cruciate tears, mid to late stage OA in the hips. Maggie was treated by Doctor David Byrd at Morehead Animal in North Carolina. Maggie presented unable to walk.
16 days post-treatment, Maggie was up walking around. 13 weeks post-treatment, you can notice Maggie's energy and mobility. Maggie enjoyed 6 years with us, pain-free, and was able to walk and run.
We then have Owen, Owen is a German Shepherd mix presented for limited mobility. He was struggling to walk from joint pain in his left leg. Owen was treated at Gold Coast Mobile by Doctor Soraya.
One month post-injection, Owen has increased mobility. Owen not only started to walk with ease again, but was even able to run. We have Sadie.
Sadie is a 4.5 year old cockapoo suffering from atopic dermatitis. She was treated by Dr.
Mike Hutchinson at Animal General. She is shown on day 0 on the far left side, the owners had exhausted all options with no relief. Doctor Mike administered an IV dose of stem cells and PRP on day 0.
Cells were banked on it as all cases should do. And at day 30, Sadie was treated again with her bank cells, no surgery needed. An improvement is already noted at the 1 month time point.
Hair is starting to grow back, redness is reduced dramatically, retreated again at day 60, again with the bank cells. Has completely grown back, redness gone, all other treatments have been stopped. Sadie is happy and comfortable.
We have another atopic dermatitis case. This dog was treated at Long Lake Animal Hospital with Doctor Lacrosse in 2020. Like Sadie, this dog had exhausted all options with no relief.
They used the same protocol as Sadie. It was that big IV dose at day zero, Bank sells for treatment at day 30 and day 60, and that bottom photo was taken on day 30, and honestly, it looks like a different dog. And that leads me to when my dog blew out her ACL one day, came in limping and I was like, All right, now what do I do?
So it was a real a moment of truth. What do I want to do? Do I do surgery on her and do the injections, or do I just do injections and I opted to do just injections.
She within days and then very quickly weeks and now months later is racing around her farm and to be honest, I can't remember which leg. We have another case with Doctor Soraya, but this was her own dog, Lera, who had a CCL tear. Doctor Saraya opted to treat with stem cells and PRP compared to doing a surgery.
And as you see in 9 days post-treatment, Lyra is already moving with ease. You see her running around, maybe a little too soon. We definitely need to send Doctor Soraya that rehab protocol following stem cell therapy.
But months later, you see her running around, clearly not bothered by the injury. Lyra actually tore her other CCL about 6 months after the initial injection, but luckily, Doctor Saraya chose to bank cells for Lyra, as I strongly recommend with every single case. Because of this decision, Dr.
Saraya was able to simply recall a vial of cells from the cryo tanks and treat Lyra immediately. No surgery required. Lera had a full recovery as well from that 2nd CCL tear and is currently living her best life.
We have Dylan. Dylan is a 5 year old German Shepherd suffering from IBD. In March 2020, he presented all the way on the left.
He was weighing only 40 pounds. He had been treated by multiple veterinarians with no improvement. All conventional therapies had failed Dylan.
He was treated with an IV dose at days 0, 30, 60, and 90. And as you can see, one year later, Dylan was up to 71 pounds. He was thriving with his family, and then in April 2022, he's still experiencing that improved quality of life and maintaining a healthy 72 pounds.
Our last case will be talked through with you by Doctor Ron Walton. Maya is an 18 month old pit bull presented for acute onset, generalised lower motor nerve weakness, and severe regurgitation. I'll let Doctor Walton walk you through Maya's case.
Case that has been one of my greatest, I feel utilisation of stem cells was the treatment of a patient with myasthenia gravis. We have very few, and if you've ever treated this devastating neuromuscular disease, you know that it can be very, very difficult both in the fact of inability to ambulate, inability to urinate, issues with mega oesophagus and these animals suffer. I ran across this article just browsing through its neuromuscular disorders, doing some research on this, and found that they had this patient that they treated with stem cells, and the patient stayed in remission for 65 months.
Now, I was kind of curious about this because I had a stem cell, I had a myasthenia gravis case, and thought, let's try this and see if it works. So just remember that myasthenia gravis is a Action against the acetylcholine antibody receptor at the neuromuscular inflate. So Mao was an 18 month old pit bull that came to me for acute onset of generalised lower motor nerve weakness and severe regurgitation.
She had a positive at 7.9 nans per litre acetylcholine receptor antibody, and if you're familiar with this disease, this is a hugely high titer. We did a tensilon test on her adrahonium, injected IV.
She went from laying recumbent, couldn't stand or walk, to jump up and ran from the back of the hospital to the front of the hospital and then promptly collapsed. Thoracic radiographs noted me oesophagus, she had no evidence of thymus or thymoma. This patient had 0 to absolutely minimal response to both pyridostigmine, prednisone, and mycophenolate.
This is a picture of Maya on presentation. So Maya is recumbent, she cannot easily stand. She can barely raise herself to a lossing point, as you'll see in this video, and is very, very debilitated.
The owner wanted to have her spayed so that she could not be bred by their other dog. So I was able to get a fat sample from the referring veterinarian. We got a very small sample, barely 20 grammes.
They brought me mostly mesenteric fat rather than alciform fat, but we were able to get a fairly good dose of 88 million stem cells that were administered. She saw a dramatic response. Within the first two weeks, she had greatly decreased regurgitation, could stand and walk on her own.
We had 19 million cells left, which were from a cryopreserve sample. We administered those at 30 days, and she continued to show improvement but was not quite normal at this stage. We had No further stem cells at this point and we had made such dramatic progress that we decided to do a second collection on this patient, and we estimated that we got about 1.17 billion cells on this, and she has received two subsequent injections, approximately 3 months apart of 136 million and 88 million stem cells.
We still have 6. Additional stem cell doses in storage for her. She is now completely normal, and over the course of this last two years, she has had a decreasing tighter on her acetylcholine esterase.
Acetylcholine receptor antibodies, and now she is actually negative on those and her megaesophagus has completely resolved. This is a picture in a video of Maya, . Last winter, a year ago, basically shows that Maya is running and able to grab this jump in the air and grab this frisbee, pretty dramatic change in this patient.
The next Video was taken this summer. This was taken just a couple of months ago this summer, and this is a video of of Maya running along the side of her owner's four wheeler. She has no exercise intolerance and it's completely normal.
And if you look at the pictures in the first video to this one, you can see how Well muscled she is at this stage. This is just an absolute dramatic response and we're at the 3-year mark right now coming up on the 3 year mark for her to be completely controlled of her myasthenia gravis. Real quick before we get to the questions, I just want to walk you through Ardent's processing of tissue for cells and speak to just how easy the process really is.
We have two, we have two options. The first is for veterinarians who bring the equipment in-house. The consumable kit is purchased, this can be processed and administered the same day in the clinic.
The dog will come in and have surgery in the morning, you collect the tissue. It takes about 30 minutes or less, open to close. Ideal collection site is again that falciform ligament, the nice fat pad in the abdomen.
It's a small incision, minimally invasive. I know some people feel weary about going into the abdomen. .
So collecting subscapular is also fun. The fat tissues handed off to your technician for processing and it takes roughly about 3 hours. There's some breaks and some machine time in there, but once the cells are isolated, they're good to be injected back in the early to late afternoon.
Leftover cells are dropped off at a UPS with a prepaid label to be sent overnight to the lab for us to bank for future use. The second option is for vets without our equipment. They follow a very similar approach, they remove the fat, but it's been sent next day air, again, a prepaid UPS label, and it comes to our labs, we do all the work for you, and it's returned in 48 hours or more, if you choose, from the date of collection, and it's ready to be injected.
Again, we make this process of bringing regenerative medicine into your practise very simple, no matter what option you choose. Keep in mind you can also participate in our proactive banking option where you collect the adipose tissue at the time of a spay or neuter, the tissues sent off and those cells are stored for the lifetime of the pet until you need it. Technically, this can be done with any surgery, so keep that in mind for your dentals as well, especially if you know the dog may need it in the future.
I'm gonna let Josh and our team discuss how you can get this started in your own practise. And with that, I want to thank everyone for taking the time out of their day to let me share a little bit about Ardent and regenerative medicine. I'm gonna let Josh briefly discuss what Arent can help with while we open the floor up to questions.
Please just type those questions in the Q&A box and we'll get started. Excellent presentation, Doctor Wright, thank you so much to our attendees, thank you for joining us. We look forward to exploring our services with your practises further.
As you think of questions to ask, I wanted to quickly highlight the options in offering our services to your pet families. As Doctor Wright mentioned, we do offer both in clinic or lab service solutions. So, whether that's equipment or using our our lab services back and forth here in the United States and into Canada, we can support you with that.
As part of the survey you take for the CE credit, you can request an equipment proposal or pricing to learn more on those different options, regardless of which option you may choose or may work best for your practise, we will support your practise by connecting you with our technical services, veterinarians, provide you. Protocol guide outlines and help with both staff education and and and ultimately pet owner education. So I'm very excited to explore these options further with all of you that joined us live and and on the recording.
Looks like we have several questions to answer, so I will turn things over back to Doctor Wright and Isabella, but excellent presentation. Thanks, Josh. Our first question is, does PRP work if just plasma and no chemical or mechanical activation, and what is the success rate of just plasma?
So, just plasma has been used in several areas. I, it is difficult because you're not actually able to extract those growth factors and get that into the body where they need it. So you are dependent upon the body to lice those platelets so that you can actually get the beneficial component, which is the growth factors.
I've seen a few studies where they just use the plasma. The results are pretty lacklustre, to be honest. So, you definitely want to be activating your, your platelets and getting those growth factors out because that's what you want and not the, the actual platelet.
Awesome. Next question is just want a guide about the most appropriate way to prepare the stem cells. Yeah, of course.
So, our team has several protocol guides that we can help you with. So based on what you're treating or how you wanna do. If you don't do it in clinic, or if then we would have a protocol, you get an on-site training, or if you want to just send it to us and let us handle it, we can do that too.
So you just reach out to Josh. All the contact information is down here. Anyone on our team would be more than happy to help.
Awesome. Next question is, can I know how the fat is processed to extract the stem cells? Yeah, so like I said, if you bring this into your clinic, your staff will actually be trained on doing this with a single-use consumable kit for each pet.
If you don't want to bring this in-house, but you still want to utilise this service, we do have an in-clinic option. I mean, yes, a lab services option where you can just take the fat tissue, send it to us, let my team handle it. I have a huge lab here where they're more than happy to process all of that for you and it comes back ready to inject.
Awesome. Is there any difference in IV versus IP injections for adiine or IBD treatments with either stem cell or PRP? Claire, that is a great question.
So, our technical services veterinarian, Doctor Larry Snyder, actually has been staying on top of this, there's a lot of literature out there to suggest right now that an IP injection for those autoimmune disorders is going to be much more beneficial than an IV dose. We know that with the IV dose, most of the cells are going to the lungs, and then they actually get trapped in the microvasculature of the lung. So by putting the IP injections in there, there's rationale that the cells are actually getting to where they need to be and can have a more targeted immune benefit.
So reach out to us, we can get some papers to you. We would love to discuss that further. Next question, how do you see this therapy fitting in to help that's deliver a spectrum of care?
That's also a really great question. So, you know, Doctor Darryl Millis is going to publish a paper saying that it comes out later this year that they found radiographically 60% of large breed dogs that appeared normal actually had radiographic signs of arthritis. For those dogs, being able to fit this proactive banking into your wellness plans and into your spectrum of care is going to be life-changing.
Not only do you get to grow the amount of pets that you have cells banked for so that when they need it, you also are gonna look at some client retention, and you know that they're gonna need it later on in life. But regenerative medicine, both stem cell and PRP are excellent tools in your toolbox for when those standard of care. Treatments that you go to have failed.
So a lot of times we see this as a last resort. Awesome. Next question is for the myasthenia gravis patient was the injection IP.
I think so IV only. This was before Snyder. So this, this one I just had to clarify, this one was IV only so we didn't have a lot of the literature and guidance that we have now about that the IP injections.
I know IP injections scare a lot of people, but yeah, that was IV only for Maya. Next question is, what countries do you work in? So, for the lab services process, it's gonna be easiest for us to do the United States and Canada, for those international countries, you are still able to get the equipment and the consumables.
We have several countries that work with us. And we are able to consent to send those consumable kits over to you. You still do get the on-site training, you still do get the support from us, the case guidance, all of the things that you would normally get if you were located in the United States or Canada.
In patients with Adi, it was mentioned that there were 2 to 3 sets of treatments on a monthly basis, or further maintenance injections needed? That's a good question. So the, the current protocol is to do that 0, 30, 60, and sometimes a 90 day re-inject as we see in those two cases that I presented, and that's gonna be based.
On the animal, but for some of the cases, they actually don't need the maintenance injections and then we've had a couple that come back every year or two and go ahead and just get that maintenance. So that's really speaking to the importance of those banks cells. Yeah.
All right, next question is, can you give us more information about what happened or what process happens in the ardent machinery? Yeah, so that's actually a super easy process. Like I said, we have a standard protocol that we use and we have the equipment.
So if you want to reach out to Josh and our team here, they can walk you through what's actually happening when you bring that equipment into your clinic. Next question is, any updates for feline renal disease. So there is a strong body of literature out there to support feline renal disease, and it is something that our veterinarians have been reaching for as a last resort for probably over a decade now.
But this is an excellent case to take to Doctor Larry Snyder because he has several out there that he can reference, . What they're really seeing is I think just improved quality of life, the numbers can sometimes fluctuate, but yeah, we do see some positive and encouraging results from that. And I would like to try that with IP.
See, we have several more questions coming in. And OK, you can answer that one. So this is a pretty interesting question that just came in.
We have tested the cell surface markers in small animals, so I do have a couple of publications on that, and they are quite different than what you see in humans, mainly because of antibody specificity. And then we see the SVF cell surface markers are different at later passages. That is correct, but what we are using is primary isolate only.
We do not culture the cells. We can absolutely send you the link for the studies after if you wanna just email us something. Mhm.
Right, I'm just gonna pop open the chat here cause I see some questions going in there. Wow. All right.
Let's scroll through these. Give us one second. And we got another couple of minutes, but if we didn't get to your question, please send us an email or give us a call.
We'd be more than happy to answer it. I see you want any side effects of PRP and MSC therapy. We have not had any reported adverse events.
I do have to report those and we currently do not have any adverse events to the PRP or the MSC therapy. See, we have a couple more. Is there?
OK, I think that we got one more. Yes, I did say that you can store PRP at the clinic, so PRP can be stored frozen in a standard freezer. Obviously, do not use one where food is stored, use one that you would utilise for other prescriptions, drugs, medications, and it can be stored frozen for up to 90 days with a single freezepa cycle.
And I see some more questions coming in about dosing. Unfortunately, we have reached the end of the hour, so if you, if we were not able to get to your question, please shoot us an email, give us a phone call, we'd be more than happy to answer your question or put you in contact with Doctor Larry Snyder. And with that, I want to thank everyone for their questions and their attendance.
This was wonderful. Thank you.
