Faculty Bibliography

BACKGROUND:Cutaneous wounds in patients with diabetes exhibit impaired healing due to physiological impediments and conventional care options are severely limited. Multipotent stromal cells (MSCs) have been touted as a powerful new therapy for diabetic tissue repair owing to their trophic activity and low immunogenicity. However, variations in sources and access are limiting factors for broader adaptation and study of MSC-based therapies. Amniotic fluid presents a relatively unexplored source of MSCs and one with wide availability. Here, we investigate the potential of amniotic fluid-derived multipotent stromal cells (AFMSCs) to restore molecular integrity to diabetic wounds, amend pathology and promote wound healing. METHOD/METHODS:diabetic mouse skin, and splinted them open to allow for humanized wound modeling. Immediately after wounding, we applied AFMSCs topically to the sites of injuries on diabetic mice, while media application only, defined as vehicle, served as controls. Post-treatment, we compared healing time and molecular and cellular events of AFMSC-treated, vehicle-treated, untreated diabetic, and non-diabetic wounds. A priori statistical analyses measures determined significance of the data. RESULT/RESULTS:Average time to wound closure was approximately 19 days in AFMSC-treated diabetic wounds. This was significantly lower than the vehicle-treated diabetic wounds, which required on average 27.5 days to heal (p < 0.01), and most similar to time of closure in wild type untreated wounds (an average of around 18 days). In addition, AFMSC treatment induced changes in the profiles of macrophage polarizing cytokines, resulting in a change in macrophage composition in the diabetic wound bed. We found no evidence of AFMSC engraftment or biotherapy induced immune response. CONCLUSION/CONCLUSIONS:Treatment of diabetic wounds using amniotic fluid-derived MSCs encourages cutaneous tissue repair through affecting inflammatory cell behavior in the wound site. Since vehicle-treated diabetic wounds did not demonstrate accelerated healing, we determined that AFMSCs were therapeutic through their paracrine activities. Future studies should be aimed towards validating our observations through further examination of the paracrine potential of AFMSCs. In addition, investigations concerning safety and efficacy of this therapy in clinical trials should be pursued.

Oral squamous cell carcinoma (OSCC) is one of the most painful cancers, which interferes with orofacial function including talking and eating. We report that legumain (Lgmn) cleaves protease-activated receptor-2 (PAR₂) in the acidic OSCC microenvironment to cause pain. Lgmn is a cysteine protease of late endosomes and lysosomes that can be secreted; it exhibits maximal activity in acidic environments. The role of Lgmn in PAR₂-dependent cancer pain is unknown. We studied Lgmn activation in human oral cancers and oral cancer mouse models. Lgmn was activated in OSCC patient tumors, compared to matched normal oral tissue. After intraplantar, facial or lingual injection, Lgmn evoked nociception in wild-type (WT) female mice but not in female mice lacking PAR₂ in Na_V1.8-positive neurons (Par₂Na_v1.8), nor in female mice treated with a Lgmn inhibitor, LI-1. Inoculation of an OSCC cell line caused mechanical and thermal hyperalgesia that was reversed by LI-1. Par₂Na_v1.8 and Lgmn deletion attenuated mechanical allodynia in female mice with carcinogen-induced OSCC. Lgmn caused PAR₂-dependent hyperexcitability of trigeminal neurons from WT female mice. Par₂ deletion, LI-1 and inhibitors of adenylyl cyclase or protein kinase A prevented the effects of Lgmn. Under acidified conditions, Lgmn cleaved within the extracellular N-terminus of PAR₂ at Asn³⁰↓Arg³¹, proximal to the canonical trypsin activation site. Lgmn activated PAR₂ by biased mechanisms in HEK293 cells to induce Ca²⁺ mobilization, cAMP formation and protein kinase A/D activation, but not β-arrestin recruitment or PAR₂ endocytosis. Thus, in the acidified OSCC microenvironment Lgmn activates PAR₂ by biased mechanisms that evoke cancer pain.SIGNIFICANCE STATEMENTOral squamous cell carcinoma (OSCC) is one of the most painful cancers. We report that legumain (Lgmn), which exhibits maximal activity in acidic environments, cleaves protease-activated receptor-2 (PAR₂) on neurons to produce OSCC pain. Active Lgmn was elevated in OSCC patient tumors, compared to matched normal oral tissue. Lgmn evokes pain-like behavior through PAR₂ Exposure of pain-sensing neurons to Lgmn decreased the current required to generate an action potential through PAR₂ Inhibitors of adenylyl cyclase and protein kinase A prevented the effects of Lgmn. Lgmn activated PAR₂ to induce calcium mobilization, cAMP formation and activation of protein kinase D and A, but not β-arrestin recruitment or PAR₂ endocytosis. Thus, Lgmn is a biased agonist of PAR₂ that evokes cancer pain.

We present a method for the efficient processing of contact and collision in volumetric elastic models simulated using the Projective Dynamics paradigm. Our approach enables interactive simulation of tetrahedral meshes with more than half a million elements, provided that the model satisfies two fundamental properties: the region of the model's surface that is susceptible to collision events needs to be known in advance, and the simulation degrees of freedom associated with that surface region should be limited to a small fraction (e.g. 5%) of the total simulation nodes. In such scenarios, a partial Cholesky factorization can abstract away the behaviour of the collision-safe subset of the face model into the Schur Complement matrix with respect to the collision-prone region. We demonstrate how fast and accurate updates of bilateral penalty-based collision terms can be incorporated into this representation, and solved with high efficiency on the GPU. We also demonstrate iterating a partial update of the element rotations, akin to a selective application of the local step, specifically on the smaller collision-prone region without explicitly paying the cost associated with the rest of the simulation mesh. We demonstrate efficient and robust interactive simulation in detailed models from animation and medical applications.

BACKGROUND:Gender-affirming surgery is becoming increasingly more common. Procedures including chest masculinization, breast augmentation, vaginoplasty, metoidioplasty, and phalloplasty routinely generate discarded tissue. The incidence of finding an occult malignancy or premalignant lesion in specimens from gender-affirming surgery is unknown. The authors therefore conducted a retrospective review of all transgender patients at their institution who underwent gender-affirming surgery to determine the incidence of precancerous and malignant lesions found incidentally. METHODS:A retrospective review of transgender patients who underwent gender-affirming surgery at the authors' institution between 2017 and 2018 performed by a single plastic surgeon and a single reconstructive urologic surgeon was conducted. Only transgender patients who underwent gender-affirming surgery that led to routine pathologic review of discarded tissue (mastectomy, vaginoplasty, vaginectomy as part of phalloplasty) were included. Charts were reviewed and patient demographics, duration of hormonal therapy, medical comorbidities, genetic risk factors for cancer, medications (including steroids or other immunosuppressants), pathology reports, and cancer management were recorded. RESULTS:Between 2017 and 2018, 295 transgender patients underwent gender-affirming surgery that generated discarded tissue sent for pathologic evaluation. During this period, 193 bilateral mastectomies, 94 vaginoplasties with orchiectomies, and eight vaginectomies were performed; 6.4 percent of all patients had an atypical lesion found on routine pathologic evaluation. CONCLUSIONS:Gender-affirming surgery is increasingly more common given the increase in access to care. The authors' review of routine pathologic specimens generated from gender-affirming surgery yielded a 6.4 percent rate of finding atypical lesions requiring further evaluation. The authors advocate that all specimens be sent for pathologic evaluation.

PURPOSE/OBJECTIVE:The aim of this work was to evaluate osseointegration of endosteal implants with two different surface treatments at early stages (~3 weeks) in the tibia of healthy and ovariectomized rabbits. MATERIALS AND METHODS/METHODS:The study comprised 10 adult New Zealand female rabbits (Oryctolagus cuniculus; 6 months and 3.0 ± 0.5 kg). Five animals were subjected to bilateral ovariectomy to mimic osteoporotic-like conditions, and the remaining rabbits (n = 5) served as the healthy control group. After 3 months, specimens from the ovariectomized and control groups were subject to implant placement in both tibiae, using two different types of surface treatment. A total of 36 implants were placed, n = 18 acid-etched and n = 18 anodized. After 3 weeks, euthanasia of the animals was performed, and samples were obtained for processing. Bone-to-implant contact and bone area fraction occupancy were quantified to evaluate the osseointegration parameters around the implant surface and within the thread area, respectively, and nanoindentation tests were performed to determine elastic modulus and hardness of the new bone. Both analyses were performed on the entire implant (total), as well as individually within the cortical and bone marrow cavity area. RESULTS:All animals were evaluated with no signs of infection or postoperative complications. The total bone-to-implant contact and bone area fraction occupancy results, independent of surface treatment, yielded significant differences between the ovariectomized and control groups (P = .002 and P < .001, respectively). In the marrow cavity, analyzing the surface treatments independently as a function of bone condition, the only differences detected were in the anodized treatment (P = .04). Regarding the elastic modulus, differences were detected only with the anodized implants between the ovariectomized and control groups (P = .015). CONCLUSION/CONCLUSIONS:At 3 weeks after implant placement, there were better osseointegration values of the implants in the healthy control group compared with the ovariectomized group independent of surface treatment. Also, specifically in the medullary region of the rabbit tibia, the acid-etched implants had more uniform osseointegration values in conditions of low-quality bone in comparison to the anodized implants, histomorphometrically and biomechanically.

The aim of this report is to describe the combination of Crouzon syndrome and acanthosis nigricans with fibrous dysplasia of the maxilla. The diagnosis of fibrous dysplasia was confirmed clinically and pathologically during Le Fort III osteotomy and midface advancement with distraction osteogenesis. Crouzon syndrome with acanthosis nigricans is a known syndrome with an incidence of 1:1,000,000. This is the first report in the literature of Crouzon syndrome and acanthosis nigricans combined with fibrous dysplasia. As all 3 pathologies are related to fibroblasts, they may be different manifestations of malfunction of a single molecular pathway. The detection of fibrous dysplasia in a patient with Crouzon syndrome and acanthosis nigricans is important because it may complicate midface osteotomies and fixation of the hardware on the bones during craniofacial surgery.