Sexual dimorphism of the synovial transcriptome underpins greater PTOA disease severity in male mice following joint injury.

OBJECTIVE: Osteoarthritis (OA) is a disease with sex-dependent prevalence and severity in both human and animal models. We sought to elucidate sex differences in synovitis, mechanical sensitization, structural damage, bone remodeling, and the synovial transcriptome in the anterior cruciate ligament rupture (ACLR) mouse model of post-traumatic OA (PTOA). DESIGN: Male and female 12-week-old C57/BL6J mice were randomized to Sham or noninvasive ACLR with harvests at 7d or 28d post-ACLR (n = 9 per sex in each group - Sham, 7d ACLR, 28d ACLR). Knee hyperalgesia, mechanical allodynia, and intra-articular matrix metalloproteinase (MMP) activity (via intravital imaging) were measured longitudinally. Trabecular and subchondral bone (SCB) remodeling and osteophyte formation were assessed by µCT. Histological scoring of PTOA, synovitis, and anti-MMP13 immunostaining were performed. NaV1.8-Cre;tdTomato mice were used to document localization and sprouting of nociceptors. Bulk RNA-seq of synovium in Sham, 7d, and 28d post-ACLR, and contralateral joints (n = 6 per group per sex) assessed injury-induced and sex-dependent gene expression. RESULTS: Male mice exhibited more severe joint damage at 7d and 28d and more severe synovitis at 28d, accompanied by 19% greater MMP activity, 8% lower knee hyperalgesia threshold, and 43% lower hindpaw withdrawal threshold in injured limbs compared to female injured limbs. Females had injury-induced catabolic responses in trabecular and SCB, whereas males exhibited 133% greater normalized osteophyte volume relative to females and sclerotic remodeling of trabecular and SCB. NaV1.8+ nociceptor sprouting in SCB and medial synovium was induced by injury and comparable between sexes. RNA-seq of synovium demonstrated similar injury-induced transcriptomic programs between the sexes at 7d, but only female mice exhibited a transcriptomic signature indicative of synovial inflammatory resolution by 28d, whereas males had persistent pro-inflammatory, pro-fibrotic, pro-neurogenic, and pro-angiogenic gene expression. CONCLUSION: Male mice exhibited more severe overall joint damage and pain behavior after ACLR, which was associated with persistent activation of synovial inflammatory, fibrotic, and neuroangiogenic processes, implicating persistent synovitis in driving sex differences in murine PTOA.

Evaluation of a coplanar 6a3ω configuration in the Hybrid III 50th percentile male head.

OBJECTIVES: In order to understand the mechanisms of traumatic brain injury (TBI) and develop proper safety measures, it is essential that accurate instrumentation methods are utilized. The brain injury criterion (BrIC) has been developed and validated to predict brain injuries in combination with the head injury criterion (Takhounts et al. 2011, 2013). Because the validated BrIC is heavily dependent on angular motion, the accuracy of any head instrumentation technique should be judged in part by its ability to measure angular motion. The main objective of this study was to evaluate a method of accurately measuring 6-degree-of-freedom (DOF) anthropomorphic test device (ATD) head kinematics using a coplanar 6 accelerometers and 3 angular rate sensors (6a3ω) configuration. METHODS: A coplanar 6a3ω configuration (c6a3ω) was implemented via a newly designed fixture. The c6a3ω fixture was placed at the center of gravity (CG) of a Hybrid III 50th percentile ATD (HIII 50) head. In addition, a tetrahedron fixture with 9 installed accelerometers (tNAAP) was externally mounted on the posterior surface of the HIII 50 skull cap. The c6a3ω setup also allowed for comparison to the 3a3ω configuration (i3a3ω) by subsequently treating the c6a3ω fixture as an i3a3ω fixture by only using accelerations and angular rates from select sensors. A total of 63 tests were conducted by impacting the head-neck apparatus at various high speeds and directions by a pneumatic ram. Normalized root mean square deviation (NRMSD), peak differences, and uncertainty were used for quantitative evaluation of the 3 configurations (e.g., c6a3ω, i3a3ω, and tNAAP). RESULTS: The average NRMSD and peak differences between the calculated angular accelerations were less than 5% between the tNAAP and the c6a3ω with 5.6% of uncertainty but greater than 18% for NRMSD and 20% for the peak differences between the tNAAP and i3a3ω with 58.2% uncertainty. Average NRMSD and peak differences between transformed resultant linear accelerations and gold standards (accelerations directly measured by accelerometers at the origin of tNAAP or c6a3ω fixtures) were also calculated. The c6a3ω had both NRMSD and peak differences less than 3% (uncertainty of 2.5%), and i3a3ω had NRMSD, peak values, and uncertainty on the order of 20% and higher. The tNAAP was slightly less accurate than the c6a3ω for transformed accelerations (NRMSD and peak differences <6%, uncertainty of 4.6%) and showed NRMSD and peak differences in the 7-8% range for angular velocity and rotation (uncertainty of 4.3 and 6.7%, respectively). CONCLUSIONS: The c6a3ω configuration exhibited much better accuracy for calculating angular acceleration and transformed linear acceleration than the i3a3ω configuration. The tNAAP showed slightly less accurate transformed linear acceleration than the c6a3ω and was demonstrated to have less accuracy than c6a3ω and i3a3ω for calculating angular velocity and rotation. The c6a3ω configuration could be a potential alternative to specialized NAAP ATD heads because all kinematics can be measured near the head CG, and 6a3ω instrumentation provides the most comprehensive 6DOF kinematics (i.e., accelerations, velocities, and displacements) with accuracy.

Traumatic brain injury in later life increases risk for Parkinson disease.

OBJECTIVE: Traumatic brain injury (TBI) is thought to be a risk factor for Parkinson disease (PD), but results are conflicting. Many studies do not account for confounding or reverse causation. We sought to address these concerns by quantifying risk of PD after TBI compared to non-TBI trauma (NTT; defined as fractures). METHODS: Using inpatient/emergency department (ED) International Classification of Disease, Ninth Revision code data for California hospitals from 2005-2006, we identified patients aged ≥55 years with TBI (n = 52,393) or NTT (n = 113,406) and without baseline PD or dementia who survived hospitalization. Using Kaplan-Meier estimates and Cox proportional hazards models (adjusted for age, sex, race/ethnicity, income, comorbidities, health care use, and trauma severity), we estimated risk of PD after TBI during follow-up ending in 2011. We also assessed interaction with mechanism of injury (fall vs nonfall) and effect of TBI severity (mild vs moderate/severe) and TBI frequency (1 TBI vs >1 TBI). RESULTS: TBI patients were significantly more likely to be diagnosed with PD compared to NTT patients (1.7% vs 1.1%, p < 0.001, adjusted hazard ratio [HR] = 1.44, 95% confidence interval [CI] = 1.31-1.58). Risk of PD was similar for TBI sustained via falls versus nonfalls (interaction p = 0.6). Assessment by TBI severity (mild TBI: HR = 1.24, 95% CI = 1.04-1.48; moderate/severe TBI: HR = 1.50, 95% CI = 1.35-1.66) and TBI frequency (1 TBI: HR = 1.45, 95% CI = 1.30-1.60; >1 TBI: HR = 1.87, 95% CI = 1.58-2.21) revealed a dose response. INTERPRETATION: Among patients aged ≥55 years presenting to inpatient/ED settings with trauma, TBI is associated with a 44% increased risk of developing PD over 5 to 7 years that is unlikely to be due to confounding or reverse causation.

A vascular model of Tsc1 deficiency accelerates renal tumor formation with accompanying hemangiosarcomas.

UNLABELLED: Tuberous sclerosis complex (TSC) is an autosomal disease caused by inactivating mutations in either of the tumor suppressor genes TSC1 or TSC2. TSC-associated tumor growth is present in multiple tissues and organs including brain, kidney, liver, heart, lungs, and skin. In the kidney, TSC angiomyolipomas have aberrant vascular structures with abnormal endothelial cells, suggesting a role for endothelial mTORC1 function. In the current report, a genetically engineered mouse model (GEMM) with a conditional knockout allele of Tsc1 with a Darpp32-Cre allele displayed accelerated formation of both kidney cystadenomas and paw hemangiosarcomas. All mutant mice developed hemangiosarcomas on multiple paws by 6 weeks of age. By 16 weeks of age, the average mutant hind paw was 4.0 mm in diameter, nearly double the size of control mice. Furthermore, the hemangiosarcomas and kidney cystadenomas were responsive to intraperitoneal rapamycin treatment. Immunoblotting and immunostaining for phospho-S6 (pS6) and phospho-CAD showed that the effect of rapamycin on tumor size was through inhibition of the mTOR signaling pathway. Finally, elevated VEGF mRNA levels were also observed in hemangiosarcoma specimens. Because paw hemangiosarcomas are easily detectable and scorable for size and growth, this novel mouse model enables accelerated in vivo drug testing for therapies of TSC-related tumors. IMPLICATIONS: These findings provide a strong rationale for simultaneous use of this conditional knockout mouse as an in vivo genetic model while seeking new cancer therapies for TSC-related tumors.