Scientific Abstract Presentations: Abstract Topics Session 1 - Non-CME Thursday, October 15 Thu, Oct 15 1:30-3:00 p.m. CDT

Background: Temporomandibular joint (TMJ) reconstruction is indicated for severe TMJ disease refractory to conservative treatment, and may be performed using costochondral grafting (CCG), stock alloplastic prostheses, or custom patient-specific alloplastic prostheses. While clinical outcomes have previously been described for each approach, no comparative cost-effective analysis between the three methods currently exists to inform value-based surgical care.

Methods: We performed an exploratory, model-based cost-effectiveness analysis comparing stock alloplastic reconstruction, custom alloplastic reconstruction, and CCG using publicly available data from peer-reviewed literature and public databases. A Markov state-transition model was created to simulate outcomes over a 15-year horizon from a societal perspective, incorporating direct medical costs and indirect productivity losses. Model inputs included revision probabilities, complication rates, and functional outcomes derived from systematic reviews and observational studies. Because TMJ-specific preference-based quality-of-life measures are unavailable in the current literature, utilities were estimated using proxy values informed by chronic pain literature that were explicitly labeled and tested in sensitivity analyses. Primary outcomes investigated were total costs, quality-adjusted life years (QALYs), and incremental cost-effectiveness ratios (ICERs). Uncertainty was evaluated using probabilistic sensitivity analysis (PSA) across 5,000 simulations.

Results: In the base-case societal analysis, stock alloplastic reconstruction was associated with mean costs of $46,876 and 9.50 QALYs, while custom alloplastic reconstruction incurred slightly higher costs ($52,308) with higher quality-adjusted survival (9.85). Compared with stock reconstruction, custom reconstruction yielded an incremental gain of 0.35 QALYs at an incremental cost of $5,432 (ICER $15,700/QALY). CCG was more costly and less effective than either method of alloplastic reconstruction. PSA showed custom reconstruction to be more frequently cost-effective than alternatives, favored in 53.0% of simulations at a willingness-to-pay (WTP) threshold of $50,000 per QALY, and 54.7% at $100,000 per QALY. No strategy was completely dominant, and these results were highly sensitive to implant cost, revision probability, and assumed utility differences.

Conclusion: This exploratory analysis suggests that custom alloplastic TMJ reconstruction may be cost-effective in certain scenarios but does not support universal preference for custom implants. Instead, these findings highlight the need for prospective studies capturing TMJ-specific preference-based quality-of-life measures and standardized cost data. Additionally, this analysis supports a value-based approach to implant selection that is appropriately tailored to individual patient scenarios and case-specific anticipated benefit.

PURPOSE: To compare long-term clinical, dental, speech, and cephalometric outcomes of patients with Pierre Robin Sequence (PRS) treated with mandibular distraction osteogenesis (MDO) versus non-operative management. METHODS: Patients with PRS managed either non-operatively or with MDO and followed for ≥9 years were included. Patients with syndromic diagnoses other than Stickler syndrome were excluded. Clinical outcomes assessed included orthognathic surgery, obstructive sleep apnea (OSA) symptoms, gastrostomy or tracheostomy tube use, temporomandibular joint (TMJ) disorders, occlusion, and speech outcomes using the Pittsburgh Weighted Speech Score. Dental outcomes, including missing teeth and occlusal classification, were evaluated using panoramic radiographs analyzed by expert orthodontists. Cephalometric measurements were obtained from standardized lateral cephalograms independently annotated by two trained reviewers, and final values were calculated as the average of both measurements. Age-adjusted linear mixed-effects regression models were used to evaluate long-term cephalometric differences between groups. RESULTS: Fifty-four patients met the inclusion criteria: 21 (38.9%) treated with MDO and 33 (61.1%) treated non-operatively. Mean follow-up was 11.3 years for the MDO group and 16.3 years for the non-operative group. There were no significant differences between groups in rates of orthognathic surgery (5% vs 18%, p = 0.227), clinical signs of OSA (19% vs 3%, p = 0.069), gastrostomy tube use (5% vs 3%, p = 1.00), tracheostomy (0% in both groups), TMJ disorders (33% vs 27%, p = 0.714), or speech outcomes (p = 0.496). Class II malocclusion was more common in the MDO group (67% vs 33%), approaching statistical significance (p = 0.054). Patients treated with MDO had a significantly higher prevalence of missing teeth (71.4% vs 27.3%, p = 0.004). Age-adjusted linear mixed-effects modeling demonstrated that MDO was associated with a larger gonial angle and increased mandibular plane divergence, reflected by greater Ar-Go-Gn (p = 0.037), Ar-Go-Me (p = 0.027), SN-Go-Gn (p = 0.015), SN-Go-Me (p = 0.015), Palatal–Mandibular Plane (p = 0.007), and Palatal–Go-Gn (p = 0.007) measurements. CONCLUSION: At long-term follow-up, MDO for PRS yields comparable clinical, airway, and speech outcomes to non-operative management. However, MDO is associated with a higher prevalence of missing teeth and persistent mandibular angular changes characterized by an increased gonial angle and vertical mandibular growth pattern.

PURPOSE: Microtia is a congenital malformation of the external ear requiring complex surgical reconstruction, typically using autologous costal cartilage or synthetic implants. However, these approaches can have drawbacks including donor site morbidity and mechanical stiffness uncharacteristic of native elastic cartilage. Tissue-engineered auricular cartilage represents a promising alternative, but current hydrogel-based constructs often suffer from contraction and loss of fine topographic detail over time. Our laboratory previously demonstrated that chondrocyte-seeded collagen matrices support elastic cartilage formation in vivo, while decellularized cartilage (DCM) improves mechanical stability but lacks chondrogenic potential alone. We hypothesize that combining DCM and chondrocytes within a type I collagen matrix will enhance volume and shape retention in vitro while supporting chondrocyte proliferation and new cartilage formation within the construct.

METHODS: Bovine auricular chondrocytes were isolated via enzymatic digestion, expanded to passage 3, and encapsulated at 25 million cells/mL into four construct formulations composed of 1% type I collagen mixed with varying amounts of DCM: (1) 100% collagen, (2) 70% collagen + 30% DCM, (3) 40% collagen + 60% DCM, and (4) 10% collagen + 90% DCM. DCM was prepared by zesting ovine rib cartilage into ~0.5 mm3 flakes, followed by chemical and enzymatic decellularization. Constructs were packed into 3D-printed scaffolds and incubated in standard media for 0, 1, 2, or 3 months. At each timepoint, constructs were assessed for gross volume, volume retention (normalized to baseline), and angle at the dome-base interface as a geometric surrogate for shape retention. Histological analysis evaluated cell distribution and cartilage-specific matrix deposition.

RESULTS: At baseline, construct volumes were comparable across groups. After 1, 2, and 3 months in vitro, constructs with higher DCM content exhibited significantly greater volume retention than collagen-only constructs. At 3 months, mean volume retention in the 90% DCM group was 98.96%±3.38%, compared to 67.55±2.68% in 30% DCM (p<0.0001) and 72.96%±2.31% in 0% DCM (p<0.0001). Angle measurements similarly demonstrated that higher DCM content preserved sharper dome-base curvature over time, while collagen-only constructs progressively flattened. Mean angle change from baseline to 3 months was 2.79°±2.02° in the 90% DCM group versus 14.17°±6.70° in 60% DCM (p<0.05), 15.11°±5.60° in 30% DCM (p<0.05), and 22.85°±8.67° in 0% DCM (p<0.001). Histology confirmed chondrocyte viability in all groups. Importantly, inclusion of DCM flakes did not hinder chondrocyte distribution within collagen in high-DCM constructs. There was no evidence of repopulation of DCM by chondrocytes.

CONCLUSIONS: Incorporation of decellularized cartilage improves volume and shape retention in bioengineered auricular constructs in vitro without negatively impacting chondrocyte viability. These findings support DCM as an internal rebar to resist contraction and preserve geometry. Ongoing biomechanical testing will further assess DCM effects on stiffness and elasticity. This composite matrix strategy represents a promising approach toward scalable, morphologically stable ear tissue engineering for microtia reconstruction and will next be tested in an in vivo model.

Purpose: Whether hemifacial microsomia (HFM) progresses during growth remains controversial. Prior studies are limited by cross-sectional design, two-dimensional imaging, or population-level comparisons rather than within-patient longitudinal assessment. We performed a multicenter longitudinal three-dimensional skeletal and soft tissue analysis to define how asymmetry evolves over time in untreated patients. Methods: Patients with unilateral HFM evaluated at two major craniofacial centers between January 2010 and December 2024 were retrospectively reviewed. Inclusion required at least two preoperative CT scans obtained ≥8 months apart. Skeletal asymmetry was quantified using affected-to-unaffected ratios for hemi-mandibular volume, ramus height, mandibular body length, maxillary height and depth, and angular measures including mental protuberance (MP) deviation and occlusal cant. Regional soft tissue thickness overlying the mandible was calculated using a projection-based method measuring perpendicular distances from mandibular cortical bone to the overlying soft tissue bilaterally. Mixed-effects models evaluated within-patient longitudinal change over time. Results: Fourteen patients (31 scans) met inclusion criteria. Mean age at initial imaging was 8.9 years (range 1.2–19.8), with mean follow-up of 3.9 years (range 0.8–10.7). Eleven patients (79%) had severe disease (Pruzansky–Kaban 2b/3). At baseline, substantial skeletal asymmetry was present, with mean hemi-mandibular volume ratio of 0.48 and mandibular body length ratio of 0.86. Mental protuberance deviation averaged 5.8°. Longitudinal analysis demonstrated progressive worsening of chin position. MP deviation increased by 0.30° per year (p<0.01), corresponding to approximately 3° over 10 years. Mandibular body length ratio decreased by 0.009 per year (p<0.01), indicating progressive relative shortening of the affected mandibular body. In absolute terms, the affected side grew 0.57 mm/year compared with 1.39 mm/year on the unaffected side (p=0.034). In contrast, hemi-mandibular volume ratio, ramus height ratio, maxillary height and depth ratios, and occlusal cant remained stable over time (all p>0.14), suggesting preservation of overall volumetric proportions despite positional progression. Soft tissue thickness ratio across the mandibular body decreased by 0.012 per year (p=0.017). The greatest progression occurred in posterior (−0.020/year, p=0.025) and middle body regions (−0.011/year, p=0.030), while the anterior body, which demonstrated the largest baseline deficit, did not significantly worsen over time. Conclusions: HFM demonstrates metric-dependent progression during growth. Chin deviation and mandibular body length worsen over time, whereas hemi-mandibular volume and ramus asymmetry remain stable. These findings suggest that positional asymmetry may continue despite preserved volumetric growth and support deferring definitive skeletal correction when feasible. Soft tissue asymmetry progresses primarily in posterior and middle mandibular regions, indicating that staged, region-specific soft tissue augmentation may be required during growth.

Purpose: Craniofrontonasal dysplasia (CFND) and frontonasal dysplasia (FND) are characterized by hypertelorism and nasal deformity, yet the nasal phenotype has not been systematically quantified. Objective characterization may inform operative planning, particularly in patients undergoing concurrent hypertelorism correction. We sought to define the nasal morphologic profile of CFND/FND and evaluate the influence of age, craniosynostosis, and hypertelorism severity. Methods: Unoperated patients with clinically confirmed CFND/FND and standardized frontal and lateral photographs were retrospectively reviewed from two tertiary craniofacial centers. Nasal measurements included nasal index, Goode's ratio, nasofrontal angle, nasal tip angle, and nasolabial angle. Measurements were compared with age-matched normative datasets. Inter-rater reliability was assessed using intraclass correlation coefficients. Mixed-effects models evaluated associations between nasal parameters, age, craniosynostosis subtype, and hypertelorism severity. Results: Twenty-three patients contributed 30 preoperative observations (mean age 3.6 ± 4.4 years; range infancy to 16 years). Craniosynostosis was present in 43% of patients (unicoronal 26%, bicoronal 17%). Inter-rater reliability was good to excellent across measurements (ICC 0.71–0.91). Compared with normative data, patients demonstrated persistently elevated nasal index across childhood, consistent with a broad nose (early childhood 130 vs 87, p=0.0020; 4–5 years 120 vs 73, p=0.030). Goode's ratio was also increased in older children (0.67 vs 0.48, p=0.049), indicating relative dorsal underprojection and shortened nasal length. Nasal index decreased modestly with age (β −2.4 per year, p=0.0070) but remained elevated relative to norms. Craniosynostosis was associated with a reduced nasofrontal angle (β −16, p=0.0070), particularly in unicoronal synostosis (β −21, p=0.0020). Increasing hypertelorism severity correlated with decreasing nasal index (β −0.11 per severity level, p=0.019), suggesting interaction between orbital width and nasal base proportions. Conclusions: CFND/FND is characterized by a persistently broad, shortened nose with relative dorsal underprojection. Although nasal index decreases with age, abnormal proportions remain. Craniosynostosis subtype influences dorsal angulation, and hypertelorism severity alters nasal base relationships. These findings provide quantitative guidance for nasal reconstruction, supporting dorsal lengthening and projection strategies and highlighting the importance of tailoring operative planning to synostosis subtype and overall facial proportions.

PURPOSE: Mandibular distraction osteogenesis (MDO) is the primary intervention for airway obstruction in Treacher Collins syndrome (TCS), yet long-term outcomes vary considerably. Some patients achieve durable airway independence after a single procedure, whereas others require serial interventions or remain tracheostomy-dependent. We sought to determine whether skeletal substrate (Pruzansky–Kaban [PK] condylar grade), lower airway (LA) disease, and baseline cephalometrics stratify longitudinal MDO outcomes and to propose a phenotype-based classification framework. METHODS: Twelve TCS patients were identified at a single tertiary craniofacial center; 10 met inclusion criteria with extended follow-up (median 10.3 years). Eight underwent MDO and were classified into three trajectory groups based on procedural burden and ultimate airway status: single-procedure success, serial distraction, and persistent tracheostomy. Two patients were managed without MDO. Bilateral PK grade and LA were recorded. Serial lateral cephalograms were analyzed when available (2–6 per patient) for SNA, SNB, and ANB. Fisher's exact and nonparametric tests assessed associations. RESULTS: Among 8 MDO patients, 3 achieved single-procedure success, 3 required serial distraction with eventual decannulation, and 2 remained tracheostomy-dependent. Bilateral PK 3/3 was associated with persistent tracheostomy dependence (2/2 tracheostomy-dependent PK3/3 vs 0/6 PK≤2b; p=0.036). Lower airway disease predicted need for multiple MDOs (4/4 LA+ vs 0/4 LA−; p=0.029) and was associated with ever requiring tracheostomy (5/5 LA+ vs 1/5 LA−; p=0.048). A PK × LA matrix segregated trajectories among PK ≤2b patients: PK ≤2b with LA− yielded exclusively single-procedure success (3/3), whereas PK ≤2b with LA+ yielded exclusively serial distraction (3/3). Both PK 3/3 patients were classified as persistent tracheostomy regardless of LA status. Baseline SNB did not differ across trajectory groups (p=0.946). In contrast, baseline ANB differed significantly (p=0.044). Serial distraction patients demonstrated large ANB (mean 22°) with relatively preserved SNA (mean 77°), consistent with isolated mandibular retrognathia. Single-procedure success patients showed moderate discrepancy (mean ANB 17°). Persistent tracheostomy patients exhibited low ANB (8°) not because of adequate mandibular position, but because both SNA (64°) and SNB (56°) were markedly depressed-reflecting global craniofacial hypoplasia. Longitudinally, SNA remained stable across groups (mean ΔSNA +1–5°), indicating changes were mandible-driven. Successful groups demonstrated substantial ANB reduction (approximately −8°) and comparable total SNB gains (+8° and +9° for single-procedure success and serial distraction, respectively). In contrast, persistent tracheostomy patients showed limited SNB improvement (+4°) and no ANB convergence (+1°). Serial distraction patients displayed oscillatory ANB and SNB trajectories requiring repeat intervention to maintain airway patency. Mechanistically, the two persistent tracheostomy patients differed: one demonstrated minimal cephalometric response to distraction, whereas the other achieved mandibular advancement but remained tracheostomy-dependent due to multi-level airway disease and glossoptosis. CONCLUSIONS: MDO trajectory in TCS appears to be determined by skeletal substrate and airway phenotype. Bilateral PK 3/3 was associated with poor distraction durability. Among patients with functional condyles (PK ≤2b), lower airway disease increases procedural burden but does not preclude eventual decannulation. Baseline ANB distinguishes isolated mandibular retrognathia from global craniofacial hypoplasia and provides prognostic insight. A phenotype-based PK × LA framework may enable prospective stratification and more informed surgical counseling.

Title: Prospective Evaluation of Autologous Fat Grafting to the Face with the use of VialityTM in Processing of Lipoaspirate

Date: November 2023 to December 2025

Background: Autologous fat transfer (AFT) is a minimally invasive medical procedure that includes aspirating adipose tissue from one part of the body and washing, filtering, and reinjecting the processed fat tissue into a different part of the body with the primary purpose of adding volume. Several published studies conclude that using a filter mesh to concentrate the fat by removing surgical tumescent fluid and blood improves fat graft retention (1,2,3). Furthermore, washing and filtration of autologous adipose tissue produce a fat graft with higher tissue viability (3,4).

Methods: This is a single-center, prospective study enrolling patients undergoing an aesthetic fat grafting procedure to the face. A total of 11 patients were enrolled in the study, and all patients received lipoaspirate processed with Viality. Patients were followed up on post-procedure months 1, 3, 6, and 12. Fat graft retention was evaluated by blinded reviewers using photographic assessment, and 3D imaging was then retrospectively compared with fat graft retention rates in AFT using traditional fat preparation techniques.

Objectives: The purpose of this research is to collect data on the short and long-term effects of facial fat grafting by injectable tissue replacement and regeneration in the midfacial zone, with the use of the VialityTM system.

Results: Historical data of AFT using ITR2 with traditional fat preparation techniques have demonstrated unique fat volume retention curves with an initial decline within 1-7 months post-operation, followed by an increase within the 8–19-month range. Patients saw a 31.6% fat retention rate during the initial decline, which increased to a 79.0% retention rate after 12 months. Data from AFT using ITR2 and the VialityTM, saw a similar trophic effect within 1-7 months and an increase within 8-19 months. The addition of VialityTM led to a 60.1% retention rate during the decline, which increased to a 76.3% retention rate.

Conclusion: The addition of the VialityTM system to the ITR2 protocol appears to stabilize the fat graft during the 1–7 month postoperative window. These findings suggest that a purer fat graft does not eliminate the trophic effect but substantially reduces the initial volume decline. Long-term retention at 12 months remained consistent with historical data, demonstrating that while VialityTM improves early retention, it does not affect final fat retention.

References:

1. Ansorge H, Garza JR, McCormack MC, et al. Autologous fat processing via the Revolve system: quality and quantity of fat retention evaluated in an animal model… doi:10.1177/1090820X14524416.

2. Salinas HM, Broelsch GF, Fernandes JR, et al. Comparative analysis of processing methods in fat grafting…doi:10.1097/PRS.0000000000000524.

3. Girard A-C, Mirbeau S, Gence L, et al. Effect of washes and centrifugation on the efficacy of lipofilling with or without local anesthetic… doi:10.1097/GOX.0000000000000465.

4. Zhu M, Zhou Z, Chen Y, et al. Supplementation of fat grafts with adipose-derived regenerative cells improves long-term graft retention… doi:10.1097/PRS.0b013e3181de1c6c.

Introduction: 2-octyl cyanoacrylate (2-OCA), used in topical skin adhesives for wound closure, is increasingly recognized as a cause of allergic contact dermatitis (ACD), producing erythema, irritation, and impaired wound healing.(1,2) Reported rates vary widely, and the population-level incidence remains uncertain as postoperative reactions are frequently unrecognized and therefore underreported.(3) We performed a systematic review and meta-analysis to characterize the incidence, clinical features, and risk factors of 2-OCA-associated ACD across surgical specialties.

Methods: We conducted a PRISMA systematic review of randomized, observational, and case-report studies in PubMed, Embase, and Web of Science (January 1, 2008, to November 7, 2025) describing human wound closure with 2-OCA-based topical skin adhesives and reporting cutaneous hypersensitivity or ACD. Risk of bias was assessed using ROBINS-I and RoB 2. For analytic cohorts reporting both exposed patients and ACD events, incidence was pooled using a random-effects inverse-variance model with prespecified subgroup analyses by surgical specialty; case reports and case series were summarized descriptively.

Results: Seventy-four studies involving 26,330 exposed patients were included; 20 analytic cohorts (25,442 patients) contributed to meta-analysis. The pooled incidence of ACD was 4% (95% CI, 3-5%) with substantial heterogeneity (I2 = 94.5%). Across analytic cohorts, study-level incidence ranged from 0.5% to 24%. Subgroup meta-analysis demonstrated pooled ACD incidence of 4% (95% CI, 2-5%) in orthopedic cohorts (13 studies, n = 24,166) and 8% (95% CI, 3-16%) in plastic-surgery cohorts (5 studies, n = 511), with lower pooled rates in dermatology and obstetrics-gynecology cohorts. Re-exposure was consistently associated with higher reaction rates, including increases from approximately 1% after initial exposure to 22% after second exposure in staged bilateral procedures, and from 12% at first exposure to 21% with repeat exposure in pediatric cohorts. Prior adhesive or contact allergy and prior cosmetic acrylate exposure were repeatedly described as contributors to risk, including sensitization from artificial nail and eyelash adhesives and other consumer acrylate-containing products.(4) Diagnosis was most commonly clinical; patch testing was used in selected cases. Treatment typically involved adhesive removal and topical corticosteroids, often with oral antihistamines; systemic corticosteroids and/or antibiotics were generally reserved for more extensive reactions or when secondary infection was suspected.

Conclusion: Allergic contact dermatitis to 2-octyl cyanoacrylate topical skin adhesives is a recurrent and clinically important complication of wound closure across multiple surgical specialties. Incidence varies substantially across study settings, with higher pooled rates observed in plastic-surgery cohorts. Risk increases markedly with re-exposure and prior sensitization, reflecting the cumulative sensitization potential of repeated 2-OCA exposure. Given the consistency of reactions reported across specialties, procedures, and adhesive formulations, 2-OCA-associated ACD should be recognized as an inherent risk of topical cyanoacrylates used in wound closure.

References: 1. Park YH, Choi JS, Choi JW, et al. Sci Rep. 2021;11:23762. doi:10.1038/s41598-021-03319-3 2. Chalmers BP, Melugin HP, Sculco PK, et al. J Arthroplasty. 2017;32:3742-3747. doi:10.1016/j.arth.2017.07.012 3. McNichol L, Lund C, Rosen T, et al. J Dermatol Nurses Assoc. 2013;5:323-338. doi:10.1097/JDN.0000000000000009 4. Gardeen S, Hylwa S. Int J Womens Dermatol. 2020;6:263-267. doi:10.1016/j.ijwd.2020.04.007

PURPOSE: Three-dimensional (3D) in vitro platforms offer powerful tools to study tumor cell behavior in physiologically relevant environments. Our laboratory previously developed a 3D biomimetic breast cancer platform demonstrating that matrix components alter tumor response to chemotherapy. Building on this work, we now apply a collagen-based 3D platform to investigate how estradiol (E2) influences proliferation and migration in both hormone receptor-positive (MCF-7) and triple-negative (MDA-MB-231) breast cancer cells, and how 3D matrix stiffness modulates these hormone-driven behaviors.

METHODS: MCF-7 and MDA-MB-231 cells expressing red fluorescent protein were exposed to graded E2 concentrations (0, 10⁻¹², 10⁻¹⁰, 10⁻⁸, 10⁻⁶, 10⁻⁴ M) in both 2D and 3D culture. In 2D, 2,000 cells were seeded per well and imaged using confocal fluorescent microscopy on Day 1 to establish baseline cell counts. Cells were cultured with E2-containing media and re-imaged on Day 5 to calculate normalized cell counts (Day 5 / Day 1). In 3D, 60,000 cells were embedded in ~50 µL constructs of 0.3% type I collagen, imaged on Day 1, and cultured in E2-containing media. On Day 10, cell counts and vertical migration were quantified using an Imaris™ machine-learning algorithm that reconstructed a regression plane and defined "migratory" cells as those located one standard deviation above or below the tumor layer's mean z-position. To assess matrix mechanics, additional cell-embedded 3D constructs were fabricated using 0.3% or 0.6% collagen and treated with either 10⁻⁶ or 0 M E2, with outcomes assessed on Day 10.

RESULTS: In 2D at 5 days, MCF-7 cells demonstrated a significant dose-dependent increase in normalized cell counts, peaking at 10⁻¹⁰ M E2 (p<0.001), whereas MDA-MB-231 exhibited stable proliferation across 0 to 10-6 M, with marked reduction at 10⁻⁴ M (p<0.001). In 3D at 10 days, MDA-MB-231 cells again maintained stable proliferation across low and intermediate doses with suppression at high doses (p<0.0001). In contrast, MCF-7 cells displayed a shifted dose-response in 3D, with maximal cell counts at 10⁻⁶ M E2 (p<0.001). Across all 3D E2 concentrations, no significant differences were observed in percentage of migratory cells or migration distance. Increased stiffness in 3D, resulting from higher collagen concentration (0.6%), suppressed cell counts (p<0.0001) and migration distance (p<0.0001) in MDA-MB-231 cells independent of E2 exposure. Meanwhile in MCF-7 cells, E2-associated increases in cell counts were most pronounced in softer 0.3% collagen and attenuated in stiffer collagen (p<0.05), without stiffness-dependent differences in migration.

CONCLUSIONS: MDA-MB-231 cells, lacking estrogen receptor (ER) expression, show minimal response to E2 in both 2D and 3D formats, with suppression at high doses likely reflecting cytotoxicity. In contrast, ER-positive MCF-7 cells demonstrate hormone-dependent proliferation, with the E2 concentration required for maximal response increasing in 3D matrices, suggesting potentially dampened hormone responsiveness in 3D environments. Matrix stiffening differentially constrains tumor behavior based on receptor status, broadly suppressing triple-negative cells, while limiting E2-associated proliferation in ER-positive cells without altering migration. These findings highlight the importance of dimensionality and matrix mechanics in shaping hormone-driven behaviors and support continued development of 3D culture models to better reflect in vivo breast cancer biology.

PURPOSE: Engineering patient-derived, three-dimensional (3D) in vitro breast cancer models holds significant potential for improving our understanding of cancer invasion within the native breast microenvironment. Our laboratory developed a tissue-engineered biomimetic breast platform incorporating patient-derived breast tissue components within a collagen matrix to recapitulate key elements of the native breast milieu. Now, we investigate tumor cell proliferation and migration behavior in plain collagen versus biomimetic (cell-containing) constructs, with specific focus on long-distance invasive behavior in two distinct breast cancer subtypes.

METHODS: Breast tissue from healthy patients undergoing reduction mammoplasty (n=6; mean age 34.5 years; mean BMI 33.99 kg/m²) was processed to isolate mature adipocytes, stromal vascular fraction, and ductal organoids, which were suspended in 0.3% type I collagen. Tri-layer 3D constructs (~50 µL total volume) were fabricated in 96-well plates and consisted of: 1) a base layer containing either triple-negative (MDA-MB-231) or estrogen receptor-positive (MCF-7) breast cancer cells tagged to red fluorescent protein, 2) a middle layer composed of either collagen alone ("Collagen") or collagen containing patient-derived breast tissue components ("Biomimetic"), and 3) a top layer containing human umbilical vascular endothelial cells. Constructs were imaged using fluorescent confocal microscopy on Day 1 to establish baseline tumor position, and again on Day 4; tumor cell counts were normalized to baseline (Day 4 / Day 1). Vertical migration was quantified using an Imaris™ machine-learning algorithm that reconstructed a regression plane and defined "migratory" cells as those located one standard deviation above or below the tumor layer's mean z-position. "Long-distance" migratory cells were defined as those traveling >100 µm above this plane. Mean migration distance among migratory cells was also calculated.

RESULTS: On Day 4, normalized tumor cell counts were comparable between Collagen and Biomimetic constructs for both MDA-MB-231 and MCF-7 cells. The overall percentage of migratory cells did not differ significantly between matrices for either subtype. In contrast, Biomimetic constructs were associated with greater invasion distance. For MDA-MB-231 cells, mean migration distance was increased (p<0.01), and the proportion of "long-distance" migratory cells (>100 µm) was significantly higher (p<0.05) in Biomimetic constructs compared with Collagen alone. For MCF-7 cells, Biomimetic constructs similarly exhibited greater mean migration distance (p<0.05) relative to Collagen controls.

CONCLUSIONS: Incorporation of patient-derived breast tissue components into this 3D platform alters breast cancer cell migration behavior. Although cell proliferation and overall proportion of migratory cells were comparable between Collagen and Biomimetic constructs, tumor cells within Biomimetic matrices exhibited a shift toward greater long-distance migratory behavior. This effect is observed not only in the highly aggressive triple-negative MDA-MB-231 breast cancer cell line but also in estrogen receptor-positive MCF-7 cells, which are traditionally considered to possess more limited invasive potential. These findings suggest that the biochemical composition of breast-derived microenvironmental components supports sustained and efficient long-distance migration, potentially uncovering invasive phenotypes not apparent in 2D or even 3D plain collagen models. Together, these data highlight the importance of incorporating patient-derived breast tissue elements into 3D culture platforms to better model clinically relevant breast cancer invasion.

Purpose: Tranexamic acid (TXA) is a hemostatic agent which has been shown to significantly reduce intraoperative blood loss. As such, it has become ubiquitous among surgeons across a variety of specialties. In the field of Plastic and Reconstructive surgery, TXA has been shown to have added effects of reducing post-operative complications such as hematoma and seroma. Despite its popularity, there remains concern among Plastic Surgeons that TXA may contribute to soft tissue ischemia by inducing microvascular thrombosis or sludging. This is of particular concern in the context of wound healing which demands a robust blood supply to adequately traffic oxygen and other growth factors to the site of injury. In this study we assessed the effect of perioperative TXA administration (either via topical or intraperitoneal [IP] routes) on wound healing in a rodent model.

Methods: Excisional wounds were created on the dorsum of thirty C57BL/6 mice using an 8mm punch biopsy. Wounds were splinted with silicone rings to minimize the effect of contracture on healing. Animals were treated with low dose TXA (10mg/kg), high dose TXA (30 mg/kg) or normal saline. Animals were randomized to receive TXA or normal saline via an IP injection or topical application directly onto the wound bed. All topical groups were mixed with 2% methylcellulose as a carrier (to prevent rapid dispersion). Gross images of each wound were taken on postoperative day (POD) 14. The rate of wound closure was assessed using planimetry compared to a baseline wound area of 8mm. The diameter of each would was also analyzed using Hematoxylin & Eosin (H&E) stained tissue specimens harvested on POD 14. CD31 immunofluorescent staining was performed in the dermal layer of each wound to assess the angiogenic response at POD 14 by quantifying the numbers of vessels per high powered field (HPF). Each of these analyses were carried out with two blinded independent raters using ImageJ.

Results: No significant differences in the rate of wound healing between groups was observed during this study (p=.45, R2 =.86) or with respect to TXA dosage (p=.67). Pooled analyses, comparing all animals treated with normal saline versus all animals treated with TXA, also demonstrated no significant difference in the rate of wound healing (p=.45). Similarly, histological analysis of wound diameter showed no significant differences between individual groups (p=.68) or during pooled analysis (p=.15). CD31 staining, likewise, revealed no significant difference in the number of vessels/HPF between groups. There were no cases of infection, hematoma, or seroma during this study.

Conclusions: These results suggest that perioperative administration TXA does not impair wound healing. Moreover, the route of delivery (local vs systemic) does not significantly impact postoperative wound healing. These data add to the increasing body of evidence that TXA should be used for its hemostatic benefits without concern regarding its effect on wound healing.

Background: Chronic wounds remain a major clinical and economic burden in the United States, affecting roughly 30 million patients and generating close to $100 billion in annual healthcare costs. Identifying affordable, biologically targeted therapies is therefore a priority. Prior work from our group demonstrated that locally injected Avena sativa-derived molecules, Avenanthramide (AVN) and β-glucan (BG), accelerate wound closure and improve scar quality, yet the mechanisms underlying these benefits remain poorly understood. This study integrates immunohistochemical and single-cell transcriptomic analyses to investigate the cellular and molecular effects of AVN and BG during wound repair.

Methods: Excisional wounds were created on the dorsal surface of C57BL/6 mice. Each mouse received subcutaneous injections of either 1% AVN, β-glucan, or PBS (control). Wounds were harvested on day 14 for histologic evaluation and single-cell RNA sequencing. Bioinformatic analyses were performed using Seurat and CellChat.

Results: AVN treatment accelerated healing and significantly altered immune cell populations. Myeloid cells comprised 42% of all cells in control wounds but only 18% in AVN treated wounds. Macrophages and neutrophils decreased from 60% and 15% in controls to 40% and 5% with AVN, respectively. In contrast, monocytes and dendritic cells increased from 4% and 5% to 10% and 20%. AVN treated macrophages had significantly elevated IGF1 expression (1.8-fold) with reduced CXCL12 expression, which is typically associated with inflammation resolution and enhanced tissue regeneration. These findings suggest that AVN promotes a transition from inflammatory to reparative immune states.

β-glucan treatment also accelerated healing but instead produced more fibroblast focused effects. Fibroblasts had the largest transcriptional shift, with a 1.5-fold increase in upregulated genes compared with control. BG treated fibroblasts upregulated Plac8, Ly6c1, Myc, and Mmp3, which suggests Wnt/β-catenin pathway involvement. Cox-2 expression was also elevated, which is consistent with activation of NF-κB mediated inflammatory signaling. Histologic and transcriptomic analyses showed higher COL7A1 expression in control compared to BG treated fibroblasts, suggesting that reduced COL7A1 may contribute to the increased scarring phenotype observed in β-glucan wounds.

Conclusion: AVN and β-glucan exert complementary yet distinct effects on wound healing. AVN reduces myeloid cell infiltration and promotes IGF1 macrophages that support inflammation resolution and regenerative repair. β-glucan drives fibroblast activation, migration, and pro-inflammatory signaling while reducing COL7A1 expression, which may explain its association with increased scarring. Thus, AVN may be beneficial in patients with chronic inflammation or impaired macrophage function (e.g., older adults), whereas β-glucan may enhance healing in immunocompromised or fibroblast deficient wounds. Future work is needed to investigate therapeutic strategies that selectively promote AVN induced immune cell recruitment and polarization or modulate BG induced fibroblast states to optimize human wound healing.

Introduction: Extearna®, identified as costal cartilage allograft specifically designed for total ear reconstruction, has emerged as a promising alternative to traditional autologous microtia reconstruction [1]. Characterization of the biomechanical and histologic properties of Extearna® versus pediatric and adolescent costal cartilage is important for translational integration. This study aims to identify the biomechanical and histological properties of Extearna® and compare them with those of autologous costal cartilage.

Methods: Twelve Extearna® and six pediatric/adolescent autologous costal cartilage (ACC) samples were obtained from patients undergoing microtia reconstruction. Samples were fixed for histology analysis and preserved for biomechanical testing. The tissues were subjected to immunohistochemistry (IHC) for collagen 2 (COL2) evaluation and stained with Safranin O (SO) and Masson's Trichrome (MT). Standardized imaging protocols were used, and images were quantified and analyzed using ImageJ. A Student's t-test was used to evaluate differences between Extearna® and ACC. Additionally, qualitative analyses were conducted to evaluate overall collagen, glycosoaminoglycan, and proteoglycan content in both Extearna® and ACC. Seven Extearna® samples underwent biomechanical testing. Mechanical property assessments included tensile values (Young's modulus, ultimate tensile strength, strain at failure, resilience, toughness) and compressive values (ultimate compressive strength). These parameters were compared with published normative biomechanical values of matched ACC.

Results: Histologically, the overall collagen fiber intensity was significantly lower in the Extearna® group (2.38 vs. 3.83, p<0.01). The proteoglycan content of the extracellular matrix showed reduced staining intensity in the Extearna® group (2.45 vs. 2.94, p<0.05). The ultimate tensile strength of Extearna® was 4.94 ± 3.16 MPa, comparable to the published normative values of 5.03 MPa [2]. Similarly, Young's modulus of Extearna® was measured at 13.54 ± 9.89 MPa, compared to the literature value of 11.1 ± 5.6 MPa [3]. Strain at failure was 0.72 ± 0.33 mm/mm. Strain-derived resilience and toughness were 1.19 ± 1.05 and 2.10 +/- 1.56 MJ m^-3, respectively. The ultimate compressive strength of Extearna® was 9.14 ± 5.28 MPa, comparable to the published normative values of 8.3 ± 1.0 MPa [4].

Discussion/Conclusion: Our study suggests that Extearna® shows reduced collagen fiber and ECM component levels compared to ACC. However, COL2 levels were similar across the tissues. The reduction in overall collagen fiber is likely a consequence of the tissue preparation process. Despite these differences, Extearna® maintains histologic similarity to ACC with respect to structural markers. Additionally, Extearna® demonstrates biomechanical performance similar to that of ACC in tensile and compressive strength prior to implantation. Future efforts include understanding the long-term clinical outcomes of Extearna® in ear reconstruction.

References: 1. https://www.mtfbiologics.org/our-products/detail/extearna-costal-cartilage-allograft 2. Alkan Z, Yigit O, Acioglu E, Bekem A, Azizli E, Kocak I, Unal A, Buyuk Y. Tensile characteristics of costal and septal cartilages used as graft materials. Arch Facial Plast Surg. 2011 Sep-Oct;13(5):322-6. doi: 10.1001/archfacial.2011.54. PMID: 21931086. 3. Weber M, Rothschild MA, Niehoff A. Anisotropic and age-dependent elastic material behavior of the human costal cartilage. Sci Rep. 2021 Jun 30;11(1):13618. doi: 10.1038/s41598-021-93176-x. PMID: 34193931; PMCID: PMC8245550. 4. Feng J, Hu T, Liu W, Zhang S, Tang Y, Chen R, Jiang X, Wei F. The biomechanical, morphologic, and histochemical properties of the costal cartilages in children with pectus excavatum. J Pediatr Surg. 2001 Dec;36(12):1770-6. doi: 10.1053/jpsu.2001.28820. PMID: 11733904.