Meiwand Bedar1, Sofia Jerez2, Nicholas Pulos3, Andre J van Wijnen4, Alexander Y Shin5. 1. Department of Orthopedic Surgery, Mayo Clinic, Rochester, MN, USA; Radboud University Medical Center, Radboud Institute for Health Sciences, Department of Plastic Surgery, Nijmegen, the Netherlands. 2. Department of Orthopedic Surgery, Mayo Clinic, Rochester, MN, USA; Department of Biochemistry and Molecular Biology, Mayo Clinic, Rochester, MN, USA. 3. Department of Orthopedic Surgery, Mayo Clinic, Rochester, MN, USA. 4. University of Vermont, Department of Biochemistry, Burlington VT, USA. 5. Department of Orthopedic Surgery, Mayo Clinic, Rochester, MN, USA. Electronic address: shin.alexander@mayo.edu.
Abstract
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.
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.
Authors: Marius Strioga; Sowmya Viswanathan; Adas Darinskas; Ondrej Slaby; Jaroslav Michalek Journal: Stem Cells Dev Date: 2012-05-09 Impact factor: 3.272
Authors: Femke Mathot; Nadia Rbia; Allen T Bishop; Steven E R Hovius; Andre J Van Wijnen; Alexander Y Shin Journal: J Plast Reconstr Aesthet Surg Date: 2019-05-22 Impact factor: 2.740
Authors: Paul J Kingham; Daniel F Kalbermatten; Daljeet Mahay; Stephanie J Armstrong; Mikael Wiberg; Giorgio Terenghi Journal: Exp Neurol Date: 2007-08-02 Impact factor: 5.330
Authors: Ruslan Masgutov; Galina Masgutova; Adelya Mullakhmetova; Margarita Zhuravleva; Anna Shulman; Alexander Rogozhin; Valeriya Syromiatnikova; Dina Andreeva; Alina Zeinalova; Kamilla Idrisova; Cinzia Allegrucci; Andrey Kiyasov; Albert Rizvanov Journal: Front Med (Lausanne) Date: 2019-04-09
Authors: Nadia Rbia; Liselotte F Bulstra; Patricia F Friedrich; Allen T Bishop; Tim H J Nijhuis; Alexander Y Shin Journal: Plast Reconstr Surg Glob Open Date: 2020-01-21