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Item QS2: Outcomes Of Pediatric Dynamic Facial Reanimation After Two Decades(Wolters Kluwer, 2021-07) Zuo, Kevin J.; Heinelt, Martina; Ho, Emily; Borschel, Gregory; Zuker, Ronald; Medicine, School of MedicinePurpose: Pediatric facial paralysis has substantial functional consequences in a growing child including impaired quality of life. Microneurovascular facial reanimation is the gold standard for smile reconstruction; however, quantitative data are lacking regarding long-term outcomes, particularly beyond 10 years. The primary objective of this study was to evaluate the long-term surgical and patient-reported outcomes after dynamic reconstruction of unilateral facial paralysis in childhood. Methods: A cross-sectional study was performed of patients in our institutional facial paralysis database (1978-2008) who underwent dynamic reconstruction of unilateral facial paralysis 20 or more years ago. All patients were treated as children with a staged cross face nerve graft and free functioning muscle transfer. Frontal facial photographs in repose and maximal smile prior to surgery, within 2 years post-surgery, and at long term follow-up were analyzed using the MEEI Face-Gram software for commissure excursion. Patient-reported outcomes were obtained using the FaCE Scale for subjective facial impairment and disability, as well as the FACE-Q Satisfaction with Outcome and FACE-Q Social Function scales. Results are reported as median [IQR] and non-parametric statistical analysis was performed with alpha of 0.05. Results: Eleven patients were included with long term follow-up of 23.7 [5.6] years (6 females, 5 males; 5 congenital, 6 acquired; age at surgery 7.3 [6.3] years). For surgical quantitative measures, commissure excursion significantly improved from prior to surgery (-1.3 [7.4] mm) compared to follow up within 2 years post-surgery (7.0 [1.7] mm) (p<0.05) and from prior to surgery compared to long term follow-up (8.3 [4.9] mm) (p<0.001). There was no statistically significant difference in commissure excursion within 2 years post-surgery and at long term follow-up (p>0.05). For patient-reported outcomes, median FaCE Scale scores showed good function for social function (81/100), oral function (88/100), facial comfort (92/100), and overall score (75/100). On the FACE-Q Satisfaction with Outcome scale, 10/11 respondents somewhat agreed or definitely agreed with the statement, “I am pleased with the result.” On the FACE-Q Social Function scale, 10/11 respondents somewhat agreed or definitely agreed with the statements, “I make a good first impression” and “I feel confident when I participate in group situations.” Conclusion: Dynamic reconstruction of unilateral facial paralysis in young children improves commissure excursion that is maintained at long-term follow up. As adults, these patients report a high level of satisfaction and social functioning with their smile reconstruction.Item QS5: The Effect of Stem Cells and Local Tacrolimus on Neurite Extension(Wolters Kluwer, 2021) Saffari, Sara; Saffari, Tiam M.; Chan, Katelyn; Borschel, Gregory H.; Shin, Alexander Y.; Surgery, School of MedicinePurpose: Application of mesenchymal stem cells (MSCs) or tacrolimus (FK506), an FDA approved immunosuppressant, to nerve grafts has been a topic of interest to enhance peripheral nerve regeneration. The aim of this study was to investigate the combined effect of MSCs and local delivery of FK506 on nerve regeneration when applied to nerve autografts and decellularized nerve allografts. Methods: A three-dimensional (3D) in vitro compartmented cell culture system, validated by Tajdaran et al (2019), consisted of rat neonatal dorsal root ganglion (DRG) adjacent to rat nerve autograft or decellularized allograft. This model was used to evaluate regenerating neurites from the DRG into the peripheral nerve scaffold. Nerve autografts and decellularized allografts were augmented with (i) dynamic undifferentiated MSC seeding, (ii) local application of FK506 (100 ng/mL) or (iii) both (N=9/group). Local application was ensured by isolating the central system (i.e. DRG side) from the peripheral system (i.e. nerve graft side), where treatment was applied. After 48-hours of incubation, DRG-nerve graft constructs were collected, fixed, sectioned and stained against neurofilament-160 to measure neurite extension. CD90 staining was used to confirm stem cell characterization. Results: All grafts treated with MSCs confirmed CD90 expression. Compared to untreated autografts, neurite extension in autografts treated with FK506 and autografts treated with MSCs and FK506 combined were found superior (P<0.001 and P=0.0001, respectively), and comparable to autografts treated with MSCs (P=0.12). Compared to untreated allografts, allografts treated with FK506, and allografts treated with MSCs and FK506 were found superior (P<0.001 and P=0.0001, respectively), and allografts treated with MSCs were found comparable (P=0.09). All autograft groups were found superior compared to their respective allograft treatment group (P<0.05). Solely allografts receiving combined treatment were found superior to untreated autografts (P<0.05). Conclusion: MSCs or FK506 treatment improved neurite outgrowth and when combined, this resulted in significant synergistic neurite extension in both autografts and allografts in comparable patterns. Schematic overview of 3D compartmented cell culture system for isolated evaluation of treatment with MSCs and local FK506 in vitro. A 3.5 mm autograft or allograft with or without undifferentiated MSC seeding is attached to a DRG. DRG-nerve graft constructs are placed through a silicone isolator in the middle of a 24-wells plate to isolate the DRG from the nerve graft. FK506 containing media was added to the nerve graft side.Item VP2: Mesenchymal Stem Cells and Local Tacrolimus Delivery Synergistically Enhance Neurite Extension(Wolters Kluwer, 2022) Saffari, Tara Sara; Saffari, Tiam M.; Chan, Katelyn; Borschel, Gregory H.; Shin, Alexander Y.; Surgery, School of MedicineINTRODUCTION: Application of mesenchymal stem cells (MSCs) or tacrolimus (FK506), an FDA approved immunosuppressant, to nerve grafts has been a topic of interest to enhance peripheral nerve regeneration. The aim of this study was to investigate the combined effect of MSCs and local delivery of FK506 on nerve regeneration when applied to nerve autografts and decellularized nerve allografts. MATERIALS AND METHODS: A three-dimensional in vitro compartmented cell culture system consisted of rat neonatal dorsal root ganglion (DRG) adjacent to rat nerve autograft or decellularized allograft. This model was used to evaluate regenerating neurites from the DRG into the peripheral nerve scaffold. Nerve autografts and decellularized allografts were augmented with (i) dynamic undifferentiated MSC seeding, (ii) local application of FK506 (100 ng/mL) or (iii) both (N=9/group). Local application was ensured by isolating the central system (i.e. DRG side) from the peripheral system (i.e. nerve graft side), where treatment was applied. After 48-hours of incubation, DRG-nerve graft constructs were collected, fixed, sectioned and stained against neurofilament-160 to measure neurite extension. CD90 staining was used to confirm stem cell characterization. RESULTS: All grafts treated with MSCs confirmed CD90 expression. Compared to untreated autografts, neurite extension in autografts treated with FK506 and autografts treated with MSCs and FK506 combined were found superior (P<0.001 and P=0.0001, respectively), and comparable to autografts treated with MSCs (P=0.12). Compared to untreated allografts, allografts treated with FK506, and allografts treated with MSCs and FK506 were found superior (P<0.001 and P=0.0001, respectively), and allografts treated with MSCs were found comparable (P=0.09). All autograft groups were found superior compared to their respective allograft treatment group (P<0.05). Solely allografts receiving combined treatment were found superior to untreated autografts (P<0.05). CONCLUSION: MSCs or FK506 treatment improved neurite outgrowth and when combined, this resulted in significant synergistic neurite extension in both autografts and allografts in comparable patterns.