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Round 2 CFU-f/ml Test Results! Marrow Cellutions vs Jamshidi

The CFU-f/ml results from Round 2 are in!

We tested the Marrow Cellutions/Maxx Regen Needle on the left posterior iliac crest against a multiport Jamshidi Needle on the right posterior iliac crest (same patient) used in a unicortical, multi-depth technique.

Technique #1:   The Marrow Cellutions Needle used a 10 cc syringe and 1cc draws per aspiration, starting about 4.5cm deep in the left posterior iliac crest…..with each 1cc aspiration being about 0.4cm – 0.5cm more superficial than the previous one.

Technique #2: A Multiport Jamshidi Needle was initially placed 1.5 cm into the right posterior iliac crest.  6cc of marrow was drawn into a 10 cc syringe that held about 1 cc of heparin. The needle was then rotated 90 degrees and another 6cc of marrow was drawn into a new 10cc syringe also preloaded with 1cc of heparin. The stylus was then replaced and the needle advanced 0.5cm.  At this new depth, the aspiration process was duplicated….resulting in another 12cc of marrow + 2cc of heparin.  This was repeated again 0.5cm deeper….and again until we had a total of 48cc of bone marrow aspirate combined with 7cc of heparin to make a total of 55cc to centrifuge in the disposable

The Total Nucleated Cell Count on the Marrow Cellutions 9cc marrow aspiration product was 21.1 million/ml  for a total of 189.9 million. The CFU-f/ml on the end product was 464/ml

The Total Nucleated Cell Count on the 55cc bone marrow aspiration was 10 million/ml for a total of 550 million. The CFU-f/ml on the pre-concentrate aspiration was 200/ml

The Total Nucleated Cell Count on the 8cc bone marrow concentrate product was 40.2 million/ml for a total of 321.6 million.  The CFU-f/ml on the post concentrate end product was 1628/ml.

SUMMARY:

Round 2 was designed to test is if the Marrow Cellutions/Maxx Regen Needle could provide a higher CFU-f/ml count than a technique where more marrow is aspirated and then centrifuged to get the end product.  The aspirations were done on the same patient on opposite iliac crests.

The Marrow Cellutions Needle aspiration provided a higher TNC count for an 8ml aspiration compared to the TNC count from a multiport Jamshidi unicortical, multidepth aspiration of 55ml.  The syringe sizes were the same but the amount of marrow drawn was different per aspiration.

The CFU-f/ml count on the final Marrow Cellutions product was 464/ml.

Author’s impression:  Drawing marrow 1 cc at a time with good technique at different depths should beat drawing marrow 6 cc at a time at different depths in a unicortical technique… if everything else is held equal.  The Marrow Cellutions needle did give a higher TNC count and CFU-f/ml count than a 55cc aspiration done with the UPICA technique prior to centrifuging and concentration and RBC removal.

The ultrasound guided posterior iliac crest (UPICA) technique uses a multi-port Jamshidi and aspirations are done at progressively deeper depths with 6cc draws per 10cc syringe used.  2 draws are done at each depth.

The CFU-f/ml count on the UPICA post filtered, centrifuged and concentrated product was CFU-f/ml of 1628/ml.

The act of centrifuging and concentrating a 55 cc marrow aspiration provided significantly higher CFU-f/ml than that obtained with the 8cc marrow aspiration done with the Marrow Cellutions Needle.

The increase in CFU-f/ml obtained was 3.5X higher by drawing 55cc of marrow instead of just 8cc and then filtering and centrifuging the marrow to create a buffy coat.

The two end products may be compared on other metrics besides TNC and CFU-f/ml but I chose these two because of how bone marrow stem cell solutions are most commonly marketed to clinicians.  There may or may not be other advantages to using unfiltered bone marrow aspiration over filtered and concentrated (RBCs removed) bone marrow concentrate…but this test wasn’t designed to evaluate RBC content or growth factor or cytokine content.

Stay Tuned for Round 3!

 

Marrow Cellutions vs Jamshidi Round 1 CFU-f/ml Test Results

Recap: On one patient I used the Marrow Cellutions/Maxx Regen needle to draw a total of 9cc of marrow from the left posterior iliac crest. On the same patient I went 1 inch superior on the same crest, and using a multiport Jamshidi with the same 10cc size syringe as the MC needle, I drew 9cc of marrow in 1cc increments with needle advancement of 0.4cm between 1cc draws (trying to best match the MC needle technique)

The TNC in the MC Needle draw was 349 million total in 9cc
The TNC in the multiport Jamshidi was 229 million total in 9cc.

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CFU-f/ml Results from Round 1

The MC Needle was 892 CFU-f/ml in 9cc.

The multi-port Jamshidi was 610 CFU-f/ml in 9cc.
So in terms of a straight aspiration, the MC needle beat the multiport Jamshidi to the tune of 46% better!

The bonus test I did on the same patient then used the same “tunnel” already made and harvested with the Jamshidi to then draw 55cc of marrow from deep to superficial….that 47cc of marrow aspirate only had 192 CFU-f/ml.

BUT WAIT 🙂

After centrifuging and concentrating even that poorly harvested sample that only had 192 CFU-f/ml…the ultimate 8cc bone marrow concentrate product had a CFU-f/ml was 1,427……higher than the MC Needle by 60%!

Take home message is that (at least on this patient) drawing 47cc of bone marrow thru a previously scavenged site and then concentrating yielded better CFU-f/ml by 60% than the MC Needle which only drew 8cc and does not concentrate the end product.

Bone Marrow Aspiration Test Round 2: Marrow Cellutions Versus Aspiration And Centrifuging

In our quest to find the best in office, ultrasound guided bone marrow aspiration solution for stem cell injections for our patients, we are continually testing novel approaches and solutions to see how they perform against current standards. In our Round 1 Test of the Marrow Cellutions/Maxx Regen Needle versus a standard multi-port Jamshidi used in a unicortical multi-depth technique, we found that the MC Needle produced a 53% better TNC compared to the multiport Jamshidi used in a unicortical multi-depth technique.  The advantage of the MC Needle disappeared however once the marrow aspiration was continued to 55cc and then centrifuged to concentrate the end product.

Chris Centeno, MD, another orthobiologic clinician and researcher completed a similar analysis recently comparing the MC Needle to his technique using a Jamshidi in a multi-cortical technique (ie. 3 cortical aspiration sites) and found that the multi-cortical technique produced a higher TNC that the MC Needle TNC even before concentrating the product from the multi-cortical technique.

In this Round 2 of our testing, we compared the MC Needle against our unicortical, multi-depth technique, harvested from opposite sides of the posterior iliac crest to minimize the scavenger effect we see in Round 1.

Round 2 Method

Our patient was 69 years old.  All samples were blinded by code known to the research team but not the lab.

The Marrow Cellution/ Maxx Regen (they are the same needle) was used per company protocol to obtain 8cc of marrow to which 1 cc of heparin was added.  The marrow aspiration was not treated further prior to sending a sample to the lab or using for injection.

A standard multiport Jamshidi was used on the opposite iliac crest to obtain 48cc of marrow using 10cc syringes with draws to 7mm.  14-15 mm was drawn at each level followed by a 0.5cm farther penetration of the needle until 48cc of marrow + 7 heparin was obtained for a total of 55cc to process.  A sample of this aspirate was sent to the lab.

The aspirate was then filtered and processed thru a Celling Biosciences disposable device and a 14 minute dual spin centrifuge to create the buffy coat.  The end product was 8cc. A sample of this bone marrow concentrate was sent to the lab.

SUMMARY OF RESULTS OF ROUND 2

Result 1: The Marrow Cellutions / Maxx Regen Needle yielded a total TNC count of 169 million cells in the 9cc volume in this 69 year old patient.

The unicortical, multi-depth Jamshidi technique yielded a total TNC of 481 million cells in the pre-centrifuge 55 cc volume.

After filtering, centrifuging, and extracting the buffy coat, the total TNC in the 8cc product was 321 million in this 69 year old patient.

DISCUSSION OF ROUND 2 RESULTS

I think our lower than average TNC counts are mostly due to the patient’s age here.

The TNC count was 90% higher in the filtered and centrifuged product which is a significant difference.

The CFU-f/ml results are pending in about 10 days and we will see if they follow the TNC trend.

There are other differences in end product such as red blood cell content and, potentially, growth factor and cytokine content, so we are still not comparing equivalent products.  However, at least with the TNC and CFU-f/ml numbers we can compare the MC Needle output with at least one example (Celling Biosciences) of a centrifuged and concentrated solution.

So far the centrifuged TNC count has been higher that the MC Needle TNC count in both of my rounds and in Dr. Centeno’s two rounds.  CFU numbers are still pending for all of us I believe….stay tuned!!

Next Free PRP and Stem Cell Seminar: May 25 at 5:45p

Our next seminar on the current applications for PRP (platelet rich plasma) and stem cells in orthopedics will be Thursday, May 25th at 5:45p at our office.  Please go to the home page at dallasprpandstemcell.com for signups and more information.

There is no charge and seminars are led by practicing, board certified orthopedic surgeons who will answer all your questions!

Bone Marrow Aspiration Test Round 1: Marrow Cellutions versus Jamshidi Needle

We are trying to find the best in office, ultrasound guided solution for our patients who we treat with bone marrow derived mesenchymal stem cells.  There is one device (Marrow Cellutions / Maxx-Regen) that proposes a technique that requires no centrifugation and injects 8-10 cc of marrow aspirated directly without further filtration, concentration of the marrow elements, or removal of red blood cells.  There is alternative technique that takes the bone marrow aspiration and then centrifuges it in order to concentrate the stem cells, some growth factors and cytokines, and to remove the red blood cells.  The end product is more concentrated that the initial aspiration….typically a 60cc bone marrow aspiration leads to a 6-10 cc bone marrow concentrate after the marrow aspiration is reduced to the final product.

Round 1 method

The Marrow Cellutions needle was used first and the Jamshidi needle was used 2.5 cm cephalad on the same side.

#1:  We used the Marrow Cellution needle to obtain 9cc of marrow added to 1 cc of ACD-A.  0.5cc of this was sent to the lab in a blinded fashion for analysis.

#2: We used a regular multiport Jamshidi and a 10 cc syringe to draw to a total of 9cc plus 1cc of ACD-A,  1 cc at a time starting at 1.5cm subcortical and advancing deeper 0.5cm per 1cc draw to best match the Marrow Cellutions technique. 0.5cc was sent to the lab in a blinded fashion for analysis.

#3: We then did a bonus test…since the Jamshidi was already 4.5 cm deep in the posterior iliac crest..we used new 10cc syringes and drew to 7cc twice at each level…withdrawing 0.5mm each time until we had 55cc of marrow.  We did this in the same cortical site and thru the same Jamshidi that we had previously used to do step #2…so this marrow aspiration was purposely subpar by our standards…we expected there to be blood dilution of this marrow aspiration.  We sent 0.5cc to the lab for analysis.

#4: We then performed a 14 minute centrifuge using the Celling Biosciences protocol and disposable and the resultant product was 8cc of bone marrow concentrate. 0.5cc of this was sent to the lab for analysis.

SUMMARY OF RESULTS OF ROUND 1

Result 1:  The Marrow Cellutions/Maxx-Regen needle did concentrate better that the multiport Jamshidi with a unicortical stick and otherwise matched syringe size and extraction technique.  TNC in the Marrow Cellutions/Maxx-Regen needle was 38.8 million/ml for a total TNC of 349 million (CFU-F/ml and total pending) (posted when available).  TNC in the Jamshidi unicortical technique was 25.4 million/ml for a total TNC of 229 million (CFU-f/ml pending). This represented a 53% increase in the MC needle over the Jamshidi for an non-contrifuged product.

Result 2: The multi-level single site Jamshidi approach with 47cc of marrow aspirated (remember this was handicapped from the study design initially) yielded a TNC of 8.7 million/ml for a total TNC of 409 million.  Because we had no plan to inject 47cc of aspirate into a joint….we centrifuged it which yielded the results in point #3

Result 3: Post filtration, centrifuging, and concentrating, the TNC was 44.6 million/ml with a total of 356 million. The volume was 8cc of bone marrow concentrate.

Brief discussion:

The MC/Maxx Regen needle did appear to concentrate TNC better than a unicortical multiport Jamshidi….53% better in this white paper.

However, even the poor aspiration technique followed by centrifuge and concentration resulted in a higher total TNC than the Marrow Cellution/Maxx-Regen needle…..356 million versus 349 million.

CFU-s are pending and will be posted.  This was all done on the same patient so we assume was no variance in marrow other than a scavenger effect from the Jamshidi needle being used after the MC needle.

Clinicians will have to decide if filtering the marrow, removing RBCs, and concentrating some of the growth factors and cytokines is a clinically useful goal for patients having these procedures.  Since with current technique that process can only be done with a centrifuge based system….clinicians may be swayed by their answer to those questions.  

Maybe a new stem cell activating drug in the future?

Some smart researchers at the University of South Carolina are looking at enzymes in our blood that may be important in the healing cascade and in putting stem cells into an “activated” state.
In their study in mice, they identified an enzyme called Hepatocyte Growth Factor Activator (HGFA) that is always in the blood stream but doesn’t activate until the body sustains an injury. HGFA, once activated by injury signals the stem cells to enter an alert state. “Alert” stem cells have a greater ability to heal and repair damaged tissues. The study is interesting and maybe has identified another thing in blood (think IRAP and a2M) that could be isolated and used to promote healing in orthobiologics. HGFA is interested in that it directly “turns on” stem cells and may be one of the factors that is giving us our clinical outcomes that hasn’t been studied much yet.
https://www.ncbi.nlm.nih.gov/pubmed/28423312
http://www.worldhealth.net/news/alerting-stem-cells-hurry-and-heal/