P9. Dimethyloxaloylglycine (DMOG) preconditioning of adipose-derived mesenchymal stem cells as strategy for enhancing posterolateral lumbar fusion in a rat model

Abstract

BACKGROUND CONTEXT Transplantation of adipose-derived mesenchymal stem cells (ADSCs) has become a promising approach for bone regeneration therapies, particularly in elderly patients. However, poor cell survival in vivo after implantation due to the ischemic tissue environment has been one of the major limitations of the effectiveness of this strategy. Hypoxia Inducible Factor-1α (HIF-1α) plays an important role in angiogenic-osteogenic coupling during bone regeneration, as it is a critical mediator of the adaptive cell response to hypoxia. DMOG treatment has been demonstrated to stabilize cellular expression of HIF-1α in cells, as well as enhance osteogenic and angiogenic capacity of MSCs. By therapeutically targeting both osteogenesis and vascularization via DMOG preconditioned MSCs, this tissue-engineering approach sought to yield superior bone quality and integration, thereby increasing rates of spinal fusion. PURPOSE We aimed to demonstrate that preconditioning adipose (ADSC)-derived (AD) MSCs with DMOG prior to implantation will prolong survival in vivo and improve spinal fusion outcomes in a rat model. STUDY DESIGN/SETTING In vivo study via a rat posterolateral spinal fusion model. PATIENT SAMPLE Syngeneic 6-8 week old Lewis rats. OUTCOME MEASURES Manual palpation scoring was performed by blinded researchers as follows: 0 = non-fused; 1 = partial fusion, some motion across operative joint; and, 2 = fused, no motion across the operated joint. MicroCT images were used to assess fusion mass volume (mm^3) via ImageJ software and determine CT fusion score (0 = non-fused; 1 = unilateral fusion; 2 = bilateral fusion). METHODS ADSCs were isolated from the inguinal fat pads of syngeneic 6-8 week old Lewis rats and cultured in vitro. When passage 1 (P1) ADSCs reached approximately 80% confluency they were pre-conditioned for 24 hours with 1 ng of DMOG (Fisher Scientific). After pre-conditioning, 2 × 10^6 ADSCs were seeded onto Vitoss (Stryker) bone graft substitute scaffolds for subsequent transplantation. Posterolateral spinal fusion surgery at L4-L5 was performed on 18 female Lewis rats divided into 2 experimental groups: [1] Vitoss+ADSCs pre-conditioned with DMOG; and, [2] Vitoss+non pre-conditioned ADSCs. Fusion was evaluated eight weeks post-surgery. Manual palpation scoring evaluated the motion across the operated joint, and MicroCT images were assessed for fusion mass volume and CT fusion. RESULTS Preliminary microCT imaging data suggest that DMOG pre-conditioned ADSCs (n=12) yielded significantly higher fusion mass volumes than non-preconditioned ADSCs (n=6) (23.49 mm^3 vs. 15.39 mm^3, respectively, p=.001). DMOG pre-conditioned ADSCs showed a trend towards higher rates of fusion and manual palpation scores compared to non-conditioned ADSCs (1.16 vs. 0.50, respectively, p=.06; and 1.25 vs. 0.66, respectively, p>.05). CONCLUSIONS In our rat posterolateral fusion model, DMOG pre-conditioned ADSCs displayed an increased fusion mass volume, CT fusion rates, and manual palpation score compared to non-conditioned ADSCs. Ongoing histological studies will evaluate whether there are any differences in the quality of bone formed within the fusion masses. Future studies will also compare whether these results are also similar in bone marrow-derived MSCs. FDA DEVICE/DRUG STATUS Dimethyloxaloylglycine (Investigational/Not approved)