Biologic Orthopedics Journal

ISOLATION OF BONE MARROW MESENCHYMAL STEM CELLS EMBEDDED IN NATIVE TISSUE STROMA YIELDS ENRICHED HARVEST, IMPROVED ADHERENCE AND PROLIFERATION, AND UNIQUE SECRETOME

2023-12-22T19:52:56+00:00

Background: The field of orthobiologics traditionally utilizes cellular products, including bone-marrow aspirate concentrate (BMAC), micronized adipose tissue, and platelet preparations to address pain from degenerative processes, orthopedic injuries and medical conditions characterized by chronic inflammation and tissue degradation. For BMAC, maximizing the concentration of mesenchymal stem cells (MSCs) in a reduced volume is thought to allow for the therapeutic delivery of the cellular concentrate, secretome, and extracellular vesicles to a site of orthopedic injury or surgical repair. The extracellular matrix (ECM) within the bone marrow stroma contains collagens and proteoglycans known to regulate cell proliferation, migration, differentiation, and cell-cell communication among resident bone marrow cells. This study aimed to evaluate the cellular effects on MSC health and function when harvested to retain their native tissue stroma.

Methods: We evaluated a novel and unique processing method and device (BMAX™) to mechanically generate a purified MSC product derived from bone marrow in a nonenzymatic manner. BMAX™ products, including cells and stroma, were plated in MSC culture media and incubated for 3–14 days (P0-P1) before evaluation with flow cytometry for cell phenotyping and immunoassays for secretome profiling.

Results: The orthobiologic product containing three-dimensional stromal components can be produced in minutes using an automated bedside device requiring minimal benchtop space. We found increased MSC adherence, improved proliferative density in culture, and significantly elevated enrichment of stromal-derived MSCs versus traditional BMAC centrifugation-based preparations. Further, we demonstrate a unique secretome profile in BMAX™ versus traditional BMAC centrifugation-based preparations.

Conclusions: These qualities provide a novel and unique platform for autologous and allogeneic bone-marrow-derived therapy to better address inflammatory and destructive processes that may improve bone-marrow-derived cell therapies’ efficacy.

COMPARATIVE METHODS FOR PROCESSING PLATELET LYSATE

2024-06-04T03:00:20+00:00

Introduction: Platelets play an important role in numerous physiologic processes through the release of bioactive proteins and growth factors contained in the α granules. These proteins can be collected after physical and chemical processes stimulate their release from platelets. After filtration, this solution contains concentrated growth factors and is then referred to as platelet lysate (PL). Among the several methods used to produce PL, no method has been established as superior in producing the highest concentration of growth factors. This study evaluated six methods of producing PL including CaCl2, freeze-thaw methods at −80˚ C and at −40˚ C, ozone exposure, USb, and USp, along with an “un-activated” Platelet-rich plasma (PRP) sample served as a control. Our goal was to evaluate each sample to determine which method produced the
highest concentration of growth factors.

Methods: PRP was produced from whole blood and subsequently divided into seven samples. One sample served as a control while the other six were used to produce PL by different methods, including ultrasonic probe homogenization, ultrasonic bath homogenization, freeze-thaw at −80˚ C, freeze-thaw at −40˚ C, addition of calcium chloride, and ozone administration.

Outcome measures: The concentration of six growth factors were measured using digital enzyme-linked immunosorbent assay from the produced samples.

Results: PL produced by ultrasonic bath produced the highest concentrations of BDNF, EGF HB-EGF, PDGF-BB, and Vascular Endothelial Growth Factor (VEGF). PL produced by ultrasonic probe produced the highest concentration of IL-1 RA. The concentration of growth factors produced by the ultrasonic bath and probe methods did not differ significantly. The growth factors found in the freeze-thaw methods did not statistically differ from control. Ozone was the least effective at releasing measurable growth factors from PRP. The calcium chloride method resulted in a clotted sample which inhibited growth factor analysis.

Conclusion: The results of this study support the effectiveness of ultrasound homogenization in releasing growth factors from PRP over other current activation methods.

ADIPOSE CELLULAR INJECTION IN THE TREATMENT OF AN INTRASUBSTANCE ACHILLES TENDON TEAR

2022-02-17T20:09:54+00:00

Introduction: We describe a case report of a patient who presented with chronic right Achilles tendon pain and weakness. Without contrast, magnetic resonance imaging of the right Achilles tendon revealed significant expansion of the Achilles tendon in the traverse and anteroposterior dimension with extensive increased T2 signal consistent with large partial-thickness Achilles tendon tear. A musculoskeletal ultrasound using a linear transducer demonstrated an anechoic tendon defect measuring 0.97 cm in the longitudinal axis with a total tendon length measuring 1.53 cm, as well as a 0.9 cm defect in the transverse axis surrounded by homogenous tendon fibers consistent with a large defect involving the distal Achilles tendon proximal to the distal insertion. The patient underwent an ultrasound-guided adipose cellular procedure using micronized fat to fill in the defect and facilitate pain reduction and tissue healing.
Conclusion: Ultrasound-guided injection of micro-fragmented adipose tissue of Achilles tendon defect can result in significant improvement in pain and function

A REVIEW OF BONE MARROW ASPIRATE CONCENTRATE THERAPY IN THE TREATMENT OF KNEE OSTEOARTHRITIS

2024-06-04T02:56:21+00:00

Purpose: Bone marrow aspirate concentrate (BMAC) provides a novel therapeutic option for knee osteoarthritis. The authors aim to systematically evaluate functional and clinical outcomes after BMAC injection treatment for knee osteoarthritis.

Methods: We used articles found in PubMed and Google Scholar using the Preferred Reporting Items for Systematic Reviews and Meta-Analyses guidelines. Studies published from January 2018 through November 2023 on patients treated with a bone marrow aspirate concentrate injection with a focus on grades I-IV Kellgren-Lawrence osteoarthritis of the knee. Reports contained functional and clinical outcomes.

Results: Eleven articles were used in the extraction of data. Eight hundred seventy-six patients were injected with BMAC and 1,010 knees with osteoarthritis were included in this literature review. For studies that passed inclusion and exclusion criteria, reported outcomes included improved pain, function, and quality of life post-procedure.

Conclusion: The literature reviewed indicates that the intraarticular injection of BMAC warrants additional investigation in treating mild to severe osteoarthritis (classified under Kellgren-Lawrence I-IV). Factors such as the preparation and concentration of BMAC remain subjects of ongoing debate and scrutiny. Consequently, further research is needed to determine the feasibility and effectiveness of BMAC as a treatment modality for knee osteoarthritis.

Level of Evidence: IV.