TY - JOUR
T1 - Dynamic seeding versus microinjection of mesenchymal stem cells for acellular nerve allograft
T2 - an in vitro comparison
AU - Bedar, Meiwand
AU - Jerez, Sofia
AU - Pulos, Nicholas
AU - van Wijnen, Andre J
AU - Shin, Alexander Y.
N1 - Funding Information:
Research reported in this publication was supported by the Mayo Clinic Orthopedic Research Review Committee (ORRC) and the National Institute of Neurological Disorders and Stroke of the National Institutes of Health under Award Number RO1 NS 102360. The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Institutes of Health
Funding Information:
Research reported in this publication was supported by the Mayo Clinic Orthopedic Research Review Committee (ORRC) and the National Institute of Neurological Disorders and Stroke of the National Institutes of Health under Award Number RO1 NS 102360. The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Institutes of Health, The study was approved by our Institutional Animal Care and Use Committee (IACUC protocol A5329-20). Meiwand Bedar: conception and execution of experiments, collection, analysis and interpretation of the data, and writing – preparation of the manuscript. Joselin S. Jerez Ortega: interpretation of the data, and writing – review and editing. Nicholas A. Pulos: interpretation of the data, acquisition of funding, and writing – review and editing. Andre J. van Wijnen: interpretation of data, and writing – review and editing. Alexander Y. Shin: conception of experiments, acquisition of funding, interpretation of data, and writing – review and editing. We would like to thank James D. Postier for the artwork of Figure 1 and Patricia F. Friedrich for her assistance with the preparations of experiments.
Publisher Copyright:
© 2022 British Association of Plastic, Reconstructive and Aesthetic Surgeons
PY - 2022
Y1 - 2022
N2 - Background: Mesenchymal stem cell (MSC)-supplemented acellular nerve allografts (ANA) are a potential strategy to improve the treatment of segmental nerve defects. Prior to clinical translation, optimal cell delivery methods must be defined. While two techniques, dynamic seeding and microinjection, have been described, the seeding efficiency, cell viability, and distribution of MSCs in ANAs are yet to be compared. Methods: Sciatic nerve segments of Sprague-Dawley rats were decellularized, and MSCs were harvested from the adipose tissue of Lewis rats. Cell viability was evaluated after injection of MSCs through a 27-gauge needle at different flow rates (10, 5, and 1 µL/min). MSCs were dynamically seeded or longitudinally injected into ANAs. Cell viability, seeding efficiency, and distribution were evaluated using LIVE/DEAD and MTS assays, scanning electron microscopy, and Hoechst staining. Results: No statistically significant difference in cell viability after injection at different flow rates was seen. After cell delivery, 84.1 ± 3.7% and 87.8 ± 2.8% of MSCs remained viable in the dynamic seeding and microinjection group, respectively (p = 0.41). The seeding efficiency of microinjection (100.4%±5.6) was significantly higher than dynamic seeding (48.1%±8.6) on day 1 (p = 0.001). Dynamic seeding demonstrated a significantly more uniform cell distribution over the course of the ANA compared to microinjection (p = 0.02). Conclusion: MSCs remain viable after both dynamic seeding and microinjection in ANAs. Higher seeding efficiency was observed with microinjection, but dynamic seeding resulted in a more uniform distribution. In vivo studies are required to assess the effect on gene expression profiles and functional motor outcomes.
AB - Background: Mesenchymal stem cell (MSC)-supplemented acellular nerve allografts (ANA) are a potential strategy to improve the treatment of segmental nerve defects. Prior to clinical translation, optimal cell delivery methods must be defined. While two techniques, dynamic seeding and microinjection, have been described, the seeding efficiency, cell viability, and distribution of MSCs in ANAs are yet to be compared. Methods: Sciatic nerve segments of Sprague-Dawley rats were decellularized, and MSCs were harvested from the adipose tissue of Lewis rats. Cell viability was evaluated after injection of MSCs through a 27-gauge needle at different flow rates (10, 5, and 1 µL/min). MSCs were dynamically seeded or longitudinally injected into ANAs. Cell viability, seeding efficiency, and distribution were evaluated using LIVE/DEAD and MTS assays, scanning electron microscopy, and Hoechst staining. Results: No statistically significant difference in cell viability after injection at different flow rates was seen. After cell delivery, 84.1 ± 3.7% and 87.8 ± 2.8% of MSCs remained viable in the dynamic seeding and microinjection group, respectively (p = 0.41). The seeding efficiency of microinjection (100.4%±5.6) was significantly higher than dynamic seeding (48.1%±8.6) on day 1 (p = 0.001). Dynamic seeding demonstrated a significantly more uniform cell distribution over the course of the ANA compared to microinjection (p = 0.02). Conclusion: MSCs remain viable after both dynamic seeding and microinjection in ANAs. Higher seeding efficiency was observed with microinjection, but dynamic seeding resulted in a more uniform distribution. In vivo studies are required to assess the effect on gene expression profiles and functional motor outcomes.
KW - Cell delivery
KW - Cell injection
KW - Dynamic seeding
KW - Mesenchymal stem cells
KW - Nerve allograft
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U2 - 10.1016/j.bjps.2022.04.017
DO - 10.1016/j.bjps.2022.04.017
M3 - Article
C2 - 35570113
AN - SCOPUS:85130365814
JO - Journal of Plastic, Reconstructive and Aesthetic Surgery
JF - Journal of Plastic, Reconstructive and Aesthetic Surgery
SN - 1748-6815
ER -