Daichi Morikawa1, Lukas N Muench2, Joshua B Baldino3, Cameron Kia4, Jeremiah Johnson4, Alexander Otto5, Leo Pauzenberger6, Felix Dyrna7, Mary Beth R McCarthy4, Augustus D Mazzocca4. 1. Department of Orthopaedic Surgery, University of Connecticut, Farmington, Connecticut, U.S.A.; Department of Orthopaedic Surgery, Juntendo University, Japan. 2. Department of Orthopaedic Surgery, University of Connecticut, Farmington, Connecticut, U.S.A.; Department of Orthopaedic Sports Medicine, Technical University of Munich, Germany. 3. Department of Orthopaedic Surgery, University of Connecticut, Farmington, Connecticut, U.S.A.. Electronic address: jbaldino@uchc.edu. 4. Department of Orthopaedic Surgery, University of Connecticut, Farmington, Connecticut, U.S.A. 5. Department of Orthopaedic Surgery, University of Connecticut, Farmington, Connecticut, U.S.A.; Department of Orthopaedic Sports Medicine, Technical University of Munich, Germany; Department of Trauma, Hand and Reconstructive Surgery, University of Münster, Germany. 6. Department of Orthopaedic Surgery, University of Connecticut, Farmington, Connecticut, U.S.A.; St. Vincent Shoulder & Sports Clinic, Vienna, Austria. 7. Department of Trauma, Hand and Reconstructive Surgery, University of Münster, Germany.
Abstract
PURPOSE: To identify an effective, nonenzymatic method for maximizing the yield of subacromial bursa-derived nucleated cells for augmenting rotator cuff repair. METHODS: Subacromial bursa (minimum 0.2 g) was collected prospectively over the supraspinatus from patients (n = 7) with at least one full-thickness tendon tear undergoing arthroscopic primary rotator cuff repair. Samples were processed and analyzed prospectively using 4 different methods: (1) mechanical digestion with scissors (chopping), (2) collagenase digestion, (3) mechanical digestion with a tissue homogenizer, and (4) whole tissue with minimal manipulation. Tissue from each method were plated and cultured in a low oxygen tension, humidified incubator for 7 days. Following incubation, cellularity was assessed with nucleated cell count using a Coulter Counter. Flow cytometry was performed on the non-enzymatic method that demonstrated the greatest cell count to confirm the presence of mesenchymal stem cells (MSCs). The Kruskal-Wallis H test and post hoc Dunn's test were used for statistical analysis. RESULTS: Following incubation, mean nucleated cell counts (cells/mL) were (1) 102,681 ± 73,249 for chopping, (2) 76,190 ± 66,275 for collagenase, (3) 31,686 ± 29,234 for homogenization, and (4) 11,162 ± 4016 for whole tissue. There was no significant difference between chopping and collagenase (P = .45) or between homogenization and collagenase (P = .52). Both chopping (P = .003) and collagenase (P = .03) produced significantly more cells when compared with whole tissue. Flow cytometry confirmed the presence of MSC markers on samples processed by chopping. CONCLUSIONS: Mechanical isolation of subacromial bursa-derived cells using a chopping technique demonstrated similar nucleated cell count compared with collagenase, along with the confirmed presence of MSCs. CLINICAL RELEVANCE: This study demonstrated a nonenzymatic, mechanical method for isolating subacromial bursa-derived cells to potentially augment rotator cuff repair. Further clinical studies are required to assess its possible advent in the tendon-bone healing process.
PURPOSE: To identify an effective, nonenzymatic method for maximizing the yield of subacromial bursa-derived nucleated cells for augmenting rotator cuff repair. METHODS: Subacromial bursa (minimum 0.2 g) was collected prospectively over the supraspinatus from patients (n = 7) with at least one full-thickness tendon tear undergoing arthroscopic primary rotator cuff repair. Samples were processed and analyzed prospectively using 4 different methods: (1) mechanical digestion with scissors (chopping), (2) collagenase digestion, (3) mechanical digestion with a tissue homogenizer, and (4) whole tissue with minimal manipulation. Tissue from each method were plated and cultured in a low oxygen tension, humidified incubator for 7 days. Following incubation, cellularity was assessed with nucleated cell count using a Coulter Counter. Flow cytometry was performed on the non-enzymatic method that demonstrated the greatest cell count to confirm the presence of mesenchymal stem cells (MSCs). The Kruskal-Wallis H test and post hoc Dunn's test were used for statistical analysis. RESULTS: Following incubation, mean nucleated cell counts (cells/mL) were (1) 102,681 ± 73,249 for chopping, (2) 76,190 ± 66,275 for collagenase, (3) 31,686 ± 29,234 for homogenization, and (4) 11,162 ± 4016 for whole tissue. There was no significant difference between chopping and collagenase (P = .45) or between homogenization and collagenase (P = .52). Both chopping (P = .003) and collagenase (P = .03) produced significantly more cells when compared with whole tissue. Flow cytometry confirmed the presence of MSC markers on samples processed by chopping. CONCLUSIONS: Mechanical isolation of subacromial bursa-derived cells using a chopping technique demonstrated similar nucleated cell count compared with collagenase, along with the confirmed presence of MSCs. CLINICAL RELEVANCE: This study demonstrated a nonenzymatic, mechanical method for isolating subacromial bursa-derived cells to potentially augment rotator cuff repair. Further clinical studies are required to assess its possible advent in the tendon-bone healing process.
Authors: Daichi Morikawa; Matthew R LeVasseur; S Brandon Luczak; Michael R Mancini; Nicholas Bellas; Mary Beth R McCarthy; Mark P Cote; Daniel P Berthold; Lukas N Muench; Augustus D Mazzocca Journal: Arthrosc Sports Med Rehabil Date: 2021-05-14
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Authors: Matthew R LeVasseur; Benjamin C Hawthorne; Michael R Mancini; Mary Beth R McCarthy; Ian J Wellington; Mark P Cote; Olga Solovyova; Vincent J Williams; Augustus D Mazzocca Journal: Arthrosc Sports Med Rehabil Date: 2021-09-08
Authors: Gregory Colbath; Alison Murray; Sandra Siatkowski; Taylor Pate; Mario Krussig; Stephan Pill; Richard Hawkins; John Tokish; Jeremy Mercuri Journal: Arthroscopy Date: 2021-06-12 Impact factor: 4.772