A patterned human neural tube model using microfluidic gradients.

The human nervous system is a highly complex but organized organ. The foundation of its complexity and organization is laid down during regional patterning of the neural tube, the embryonic precursor to the human nervous system. Historically, studies of neural tube patterning have relied on animal models to uncover underlying principles. Recently, models of neurodevelopment based on human pluripotent stem cells, including neural organoids1-5 and bioengineered neural tube development models6-10, have emerged. However, such models fail to recapitulate neural patterning along both rostral-caudal and dorsal-ventral axes in a three-dimensional tubular geometry, a hallmark of neural tube development. Here we report a human pluripotent stem cell-based, microfluidic neural tube-like structure, the development of which recapitulates several crucial aspects of neural patterning in brain and spinal cord regions and along rostral-caudal and dorsal-ventral axes. This structure was utilized for studying neuronal lineage development, which revealed pre-patterning of axial identities of neural crest progenitors and functional roles of neuromesodermal progenitors and the caudal gene CDX2 in spinal cord and trunk neural crest development. We further developed dorsal-ventral patterned microfluidic forebrain-like structures with spatially segregated dorsal and ventral regions and layered apicobasal cellular organizations that mimic development of the human forebrain pallium and subpallium, respectively. Together, these microfluidics-based neurodevelopment models provide three-dimensional lumenal tissue architectures with in vivo-like spatiotemporal cell differentiation and organization, which will facilitate the study of human neurodevelopment and disease.

A Standardized Set of MoClo-Compatible Inducible Promoter Systems for Tunable Gene Expression in Yeast.

Small-molecule control of gene expression underlies the function of numerous engineered gene circuits that are capable of environmental sensing, computation, and memory. While many recently developed inducible promoters have been tailor-made for bacteria or mammalian cells, relatively few new systems have been built for Saccharomyces cerevisiae, limiting the scale of synthetic biology work that can be done in yeast. To address this, we created the yeast Tunable Expression Systems Toolkit (yTEST), which contains a set of five extensively characterized inducible promoter systems regulated by the small-molecules doxycycline (Dox), abscisic acid (ABA), danoprevir (DNV), 1-naphthaleneacetic acid (NAA), and 5-phenyl-indole-3-acetic acid (5-Ph-IAA). Assembly was made to be compatible with the modular cloning yeast toolkit (MoClo-YTK) to enhance the ease of use and provide a framework to benchmark and standardize each system. Using this approach, we built multiple systems with maximal expression levels greater than those of the strong constitutive TDH3 promoter. Furthermore, each of the five classes of systems could be induced at least 60-fold after a 6 h induction and the highest fold change observed was approximately 300. Thus, yTEST provides a reliable, diverse, and customizable set of inducible promoters to modulate gene expression in yeast for applications in synthetic biology, metabolic engineering, and basic research.

A Microfluidic Platform for Screening Gene Expression Dynamics across Yeast Strain Libraries.

The relative ease of genetic manipulation in S. cerevisiae is one of its greatest strengths as a model eukaryotic organism. Researchers have leveraged this quality of the budding yeast to study the effects of a variety of genetic perturbations, such as deletion or overexpression, in a high-throughput manner. This has been accomplished by producing a number of strain libraries that can contain hundreds or even thousands of distinct yeast strains with unique genetic alterations. While these strategies have led to enormous increases in our understanding of the functions and roles that genes play within cells, the techniques used to screen genetically modified libraries of yeast strains typically rely on plate or sequencing-based assays that make it difficult to analyze gene expression changes over time. Microfluidic devices, combined with fluorescence microscopy, can allow gene expression dynamics of different strains to be captured in a continuous culture environment; however, these approaches often have significantly lower throughput compared to traditional techniques. To address these limitations, we have developed a microfluidic platform that uses an array pinning robot to allow for up to 48 different yeast strains to be transferred onto a single device. Here, we detail a validated methodology for constructing and setting up this microfluidic device, starting with the photolithography steps for constructing the wafer, then the soft lithography steps for making polydimethylsiloxane (PDMS) microfluidic devices, and finally the robotic arraying of strains onto the device for experiments. We have applied this device for dynamic screens of a protein aggregation library; however, this methodology has the potential to enable complex and dynamic screens of yeast libraries for a wide range of applications. Key features • Major steps of this protocol require access to specialized equipment (i.e., microfabrication tools typically found in a cleanroom facility and an array pinning robot). • Construction of microfluidic devices with multiple different feature heights using photolithography and soft lithography with PDMS. • Robotic spotting of up to 48 different yeast strains onto microfluidic devices.

Anthropomorphic Characterization of Ankle Joint.

Even though total ankle replacement has emerged as an alternative treatment to arthrodesis, the long-term clinical results are unsatisfactory. Proper design of the ankle device is required to achieve successful arthroplasty results. Therefore, a quantitative knowledge of the ankle joint is necessary. In this pilot study, imaging data of 22 subjects (with both females and males and across three age groups) was used to measure the morphological parameters of the ankle joint. A total of 40 measurements were collected by creating sections in the sagittal and coronal planes for the tibia and talus. Statistical analyses were performed to compare genders, age groups, and image acquisition techniques used to generate 3D models. About 13 measurements derived for parameters (TiAL, SRTi, TaAL, SRTa, TiW, TaW, and TTL) that are very critical for the implant design showed significant differences (p-value < 0.05) between males and females. Young adults showed a significant difference (p-value < 0.05) compared to adults for 15 measurements related to critical tibial and talus parameters (TiAL, TiW, TML, TaAL, SRTa, TaW, and TTL), but no significant differences were observed between young adults and older adults, and between adults and older adults for most of the parameters. A positive correlation (r > 0.70) was observed between tibial and talar width values and between the sagittal radius values. When compared with morphological parameters obtained in this study, the sizes of current total ankle replacement devices can only fit a very limited group of people in this study. This pilot study contributes to the comprehensive understanding of the effects of gender and age group on ankle joint morphology and the relationship between tibial and talus parameters that can be used to plan and design ankle devices.

Fabrication of Microfluidic Devices for Continuously Monitoring Yeast Aging.

For several decades, aging in Saccharomyces cerevisiae has been studied in hopes of understanding its causes and identifying conserved pathways that also drive aging in multicellular eukaryotes. While the short lifespan and unicellular nature of budding yeast has allowed its aging process to be observed by dissecting mother cells away from daughter cells under a microscope, this technique does not allow continuous, high-resolution, and high-throughput studies to be performed. Here, we present a protocol for constructing microfluidic devices for studying yeast aging that are free from these limitations. Our approach uses multilayer photolithography and soft lithography with polydimethylsiloxane (PDMS) to construct microfluidic devices with distinct single-cell trapping regions as well as channels for supplying media and removing recently born daughter cells. By doing so, aging yeast cells can be imaged at scale for the entirety of their lifespans, and the dynamics of molecular processes within single cells can be simultaneously tracked using fluorescence microscopy. Key features This protocol requires access to a photolithography lab in a cleanroom facility. Photolithography process for patterning photoresist on silicon wafers with multiple different feature heights. Soft lithography process for making PDMS microfluidic devices from silicon wafer templates.

Implementation of a Multidisciplinary Preoperative Protocol for Geriatric Hip Fractures Improves Time to Surgery at a Level III Trauma Center.

Introduction: Hip fractures are common among the elderly, and delays in time to surgery (TTS) and length of stay (LOS) are known to increase mortality risk in these patients. Preoperative multidisciplinary protocols for hip fracture management are effective at larger trauma hospitals. The purpose of this study is to evaluate the effect of a similar multidisciplinary preoperative protocol for geriatric hip fracture patients at our Level III trauma center. Materials and Methods: In this single-center retrospective study, patients aged 65 and older who were admitted from March 2016 to December 2018 (pre-protocol group, Cohort #1, n = 247) and from August 2021 to September 2022 (post-protocol group, Cohort #2, n = 169) were included. Demographic information, TTS, and LOS were obtained and compared using Student's t-test and Chi-square testing. Results: There was a significant decrease in TTS in Cohort #2 compared to Cohort #1 (P < .001). There was a significant increase in LOS in Cohort #2 compared to Cohort #1 (P < .05), but when comparing a subset of Cohort #2 (Subgroup 2B, patients admitted from May to September 2022 when the effects of COVID-19 were likely dissipated) to Cohort #1, there was no significant difference in LOS (P = .13). For patients admitted to skilled nursing facilities (SNF), LOS in Cohort #2 was significantly longer than in Cohort #1 (P = .001). Discussion: In general, Level III hospitals have fewer perioperative resources compared to larger Level I hospitals. Despite this fact, this multidisciplinary preoperative protocol effectively reduced TTS which improves mortality risk in elderly patients. LOS is a multifactorial variable, and we believe the COVID-19 pandemic was a significant confounder that reduced available SNF beds in our area which prolonged the average LOS in Cohort #2. Conclusion: A multidisciplinary preoperative protocol for geriatric hip fracture management can improve efficiency of getting patients to surgery at Level III trauma centers.

Age-dependent aggregation of ribosomal RNA-binding proteins links deterioration in chromatin stability with challenges to proteostasis.

Chromatin instability and protein homeostasis (proteostasis) stress are two well-established hallmarks of aging, which have been considered largely independent of each other. Using microfluidics and single-cell imaging approaches, we observed that, during the replicative aging of Saccharomyces cerevisiae, a challenge to proteostasis occurs specifically in the fraction of cells with decreased stability within the ribosomal DNA (rDNA). A screen of 170 yeast RNA-binding proteins identified ribosomal RNA (rRNA)-binding proteins as the most enriched group that aggregate upon a decrease in rDNA stability induced by inhibition of a conserved lysine deacetylase Sir2. Further, loss of rDNA stability induces age-dependent aggregation of rRNA-binding proteins through aberrant overproduction of rRNAs. These aggregates contribute to age-induced proteostasis decline and limit cellular lifespan. Our findings reveal a mechanism underlying the interconnection between chromatin instability and proteostasis stress and highlight the importance of cell-to-cell variability in aging processes.

Patterning of brain organoids derived from human pluripotent stem cells.

The emerging technology of brain organoids deriving from human pluripotent stem cells provides unprecedented opportunities to study human brain development and associated disorders. Various brain organoid protocols have been developed that can recapitulate some key features of cell type diversity, cytoarchitectural organization, developmental processes, functions, and pathologies of the developing human brain. In this review, we focus on patterning of human stem cell-derived brain organoids. We start with an overview of general procedures to generate brain organoids. We then highlight some recently developed brain organoid protocols and chemical cues involved in modeling development of specific human brain regions, subregions, and multiple regions together. We also discuss limitations and potential future improvements of human brain organoid technology.

Evaluation of a Novel Multidisciplinary Preoperative Workup Strategy for Geriatric Hip Fractures.

OBJECTIVE: To determine the financial and clinical impact of a standardized, multidisciplinary team for surgical clearance and optimization in geriatric hip fracture patients. DESIGN: Retrospective case series. SETTING: Level-1 trauma center. PATIENTS: One hundred twenty-four geriatric patients (age >65 years old) in the preprotocol group (cohort 1; January 2017-December 2018) and 98 geriatric patients in the postprotocol group (cohort 2; October 2019-January 2021) with operative hip fractures. INTERVENTION: Implementation of a multidisciplinary team protocol consisting of Anesthesiology, Internal Medicine and Orthopedic Surgery departments for the assessment of medical readiness and optimization for surgical intervention in geriatric hip fractures. MAIN OUTCOME MEASURES: Rate of cardiology consultation, need for cardiac workup (echocardiography stress testing, heath catheterization), time to medical readiness (TTMR), time to surgery, case-cancellation rate, length of stay (LOS), and total hospitalization charges. RESULTS: Following implementation of the new protocol, there were significant ( P < 0.001) decreases in TTMR (19 vs. 11 hours), LOS (149 vs. 120 hours), case cancellation rate, and total hospital charges ($84,000 vs. $62,000). There were no significant differences with respect to in-hospital complications or readmission rates/mortality rates at 1 year. CONCLUSIONS: Following implementation of a protocolized, multidisciplinary approach to optimizing geriatric fracture patients, we were able to demonstrate a reduction in unnecessary preoperative testing, TTMR for surgery, case cancellation rate, LOS, and total hospitalization charge-without a concomitant increase in complications or mortality. This study highlights that standardization of the perioperative care for geriatric hip fracture patients can provide effective patient care while also lowering financial and logistical burden in care for these injuries. LEVEL OF EVIDENCE: Therapeutic Level III. See Instructions for Authors for a complete description of levels of evidence.

Outcome Reporting in Total Ankle Arthroplasty: A Systematic Review.

Total ankle arthroplasty is an increasingly utilized treatment for ankle arthritis due to decreasing failure rates and improved outcomes. However, the literature on this procedure remains heterogeneous with large variability in outcome reporting methods. PRISMA guidelines were used to systematically review outcome reporting measures used in prospective studies and randomized control trials on total ankle arthroplasty published in 15 high-impact journals between Jan 1, 2009-May 1, 2020. A total of 43 studies were included and outcome measures were grouped into seven categories: pain, subjective function, patient satisfaction, complications, objective function, implant survivorship, and imaging. The most common topic of study was implant design followed by differences in outcomes when comparing total ankle arthroplasty and arthrodesis. The most commonly reported outcome measure was post-surgical complications. No study reported on all seven categories, while 22 (51.2%) studies reported on four or more. Subjective measures had significant variability with fifteen different Patient Reported Outcome Measures used across the studies. While the included studies were quite comprehensive, there was little consistency in reporting outcomes after total ankle arthroplasty. With improving outcomes and techniques in total ankle arthroplasty, and thus an expected increase in utilization and number of published studies, efforts should be made to use commonly employed outcome reporting methods to facilitate comparison of results across studies.

Biomechanical Comparison of Tape Versus Suture in Simulated Achilles Tendon Midsubstance Rupture.

As sutures have progressed in strength, increasing evidence supports the suture tendon interface as the site where most tendon repairs fail. We hypothesized that suture tape would have a higher load to failure versus polyblend suture due to its larger surface area. Eleven matched pairs of cadaveric Achilles tendons were sutured with 2 mm wide braided ultrahigh molecular weight polyethylene tape (Tape) or 2 mm wide braided ultrahigh molecular weight polyethylene suture (Suture) using a Krackow repair method. All Achilles repair constructs were cyclically loaded, after which they were loaded to failure. Change in suture footprint height, clinical and ultimate load to failure, and location of failure was recorded. Clinical loads to failure for Tape and Suture were 290.4 ± 74.8 and 231.7 ± 70.4 Newtons, respectively (p= .01). Ultimate loads to failure for Tape and Suture were 352.9 ± 108.1 and 289.8 ± 53.7 Newtons, respectively (p = .11). Cyclic testing resulted in significant changes in footprint height for both Tape and Suture, but the 2 sutures did not differ in terms of the magnitude of change in footprint height (p = .52). The suture tendon interface was the most common site of failure for both Tape and Suture. Our results suggest that Tape may provide added repair strength in vivo for Achilles midsubstance rupture.

Regulatory protein HilD stimulates Salmonella Typhimurium invasiveness by promoting smooth swimming via the methyl-accepting chemotaxis protein McpC.

In the enteric pathogen Salmonella enterica serovar Typhimurium, invasion and motility are coordinated by the master regulator HilD, which induces expression of the type III secretion system 1 (T3SS1) and motility genes. Methyl-accepting chemotaxis proteins (MCPs) detect specific ligands and control the direction of the flagellar motor, promoting tumbling and changes in direction (if a repellent is detected) or smooth swimming (in the presence of an attractant). Here, we show that HilD induces smooth swimming by upregulating an uncharacterized MCP (McpC), and this is important for invasion of epithelial cells. Remarkably, in vitro assays show that McpC can suppress tumbling and increase smooth swimming in the absence of exogenous ligands. Expression of mcpC is repressed by the universal regulator H-NS, which can be displaced by HilD. Our results highlight the importance of smooth swimming for Salmonella Typhimurium invasiveness and indicate that McpC can act via a ligand-independent mechanism when incorporated into the chemotactic receptor array.

A programmable fate decision landscape underlies single-cell aging in yeast.

Chromatin instability and mitochondrial decline are conserved processes that contribute to cellular aging. Although both processes have been explored individually in the context of their distinct signaling pathways, the mechanism that determines which process dominates during aging of individual cells is unknown. We show that interactions between the chromatin silencing and mitochondrial pathways lead to an epigenetic landscape of yeast replicative aging with multiple equilibrium states that represent different types of terminal states of aging. The structure of the landscape drives single-cell differentiation toward one of these states during aging, whereby the fate is determined quite early and is insensitive to intracellular noise. Guided by a quantitative model of the aging landscape, we genetically engineered a long-lived equilibrium state characterized by an extended life span.

Biomechanical Behavior of a Variable Angle Locked Tibiotalocalcaneal Construct.

This paper examines the mechanics of the tibiotalocalcaneal construct made with a PHILOS plating system. A failed device consisting of the LCP plate and cortical, locking, and cannulated screws was used to perform the analysis. Visual, microstructure, and fractographic examinations were carried out to characterize the fracture surface topology. These examinations revealed the presence of surface scratching, inclusions, discoloration, corrosion pits, beach marks, and cleavage and striations on the fracture surface. Further examination of the material crystallography and texture revealed an interaction of S, Ni, and Mo-based inclusions that may have raised pitting susceptibility of the device made with Stainless Steel 316L. These features suggest that the device underwent damage by pitting the corrosion-fatigue mechanism and overloading towards the end to fail the plate and screws in two or more components. The screws failed via conjoint bending and torsion fatigue mechanisms. Computer simulations of variable angle locking screws were performed in this paper. The material of construction of the device was governed by ASTM F138-8 or its ISO equivalent 5832 and exhibited inconsistencies in chemistry and hardness requirements. The failure conditions were matched in finite element modeling and those boundary conditions discussed in this paper.

Improvement and Retention of Arthroscopic Skills in Novice Subjects Using Fundamentals of Arthroscopic Surgery Training (FAST) Module.

INTRODUCTION: Analysis of the Fundamentals of Arthroscopy Surgery Training (FAST) workstation regarding increased proficiency and retention of basic arthroscopy skills in novice subjects. METHODS: First-year medical students from a single allopathic medical school performed weekly standardized FAST workstation modules for a consecutive 6 weeks. Primary outcomes evaluated were time to task completion and error rate on specific modules. Scores were analyzed using a one-way repeated measures analysis of variance design for overall trends in time and errors over the 6-week study. Psychomotor retention was analyzed after a 12-week and 24-week interlude. RESULTS: Across the initial 6-week study, the average time to complete all modules at the workstation decreased significantly (P < 0.001) with a mean reduction in the total workstation time of 21.9 minutes (s = 8.12 minutes). Weekly comparisons showed the most significant improvement from week 1 to week 2 for the total workstation time (P < 0.001). Results after a 12-week and 24-week interval of inactivity demonstrated no significant difference in the mean workstation time or errors when compared with the original 6-week study. DISCUSSION: The FAST workstation significantly improved the task performance of novice participants over a 6-week period with no significant deterioration in task performance after 12 and 24 weeks of inactivity.

The Popularity of Outcome Measures Used in the Foot and Ankle Literature.

Background. Outcome measures are frequently employed in clinical studies to determine the efficacy of orthopaedic surgical procedures. However, substantial variability exists among the outcome instruments utilized in foot and ankle (F&A) literature. The purpose of this study is to determine the number of outcome measures reported in F&A literature recently published in major orthopaedic journals and the association between study characteristics and the use of particular outcome measurement categories. Methods. All manuscripts published in 6 major orthopaedic journals between 2013-2017 reporting at least one clinical outcome measure were collected. For each manuscript, the journal, title, authors, country/region of origin, level of evidence, topic, and anatomic location were recorded. Outcome measures were characterized as generic, F&A specific, and disease specific. Poisson regression with robust error variance was used to test for association between study characteristics and outcome measure categories. Results. A total of 541 F&A articles were included with fifty-two different outcome measures reported. The most popular tool was the American Orthopaedic Foot and Ankle Score (AOFAS) (56.9%). Generic outcome measures were used in 331 (61.1%) studies, while 440 (81.3%) studies used F&A specific measures and 64 (11.8%) used disease-specific measures. The use of generic and disease-specific outcome measures was associated with a higher level of evidence (p < 0.001). Conclusion. AA substantial variety of outcome measures are employed among recent published studies, with many studies utilizing non-validated measures. Reporting a combination of validated and focused outcome measures is necessary to improve the quality and generalizability of published studies in foot and ankle literature. Levels of Evidence: Level II: Systematic review.

Advances in quantitative biology methods for studying replicative aging in Saccharomyces cerevisiae.

Aging is a complex, yet pervasive phenomenon in biology. As human cells steadily succumb to the deteriorating effects of aging, so too comes a host of age-related ailments such as neurodegenerative disorders, cardiovascular disease and cancer. Therefore, elucidation of the molecular networks that drive aging is of paramount importance to human health. Progress toward this goal has been aided by studies from simple model organisms such as Saccharomyces cerevisiae. While work in budding yeast has already revealed much about the basic biology of aging as well as a number of evolutionarily conserved pathways involved in this process, recent technological advances are poised to greatly expand our knowledge of aging in this simple eukaryote. Here, we review the latest developments in microfluidics, single-cell analysis and high-throughput technologies for studying single-cell replicative aging in S. cerevisiae. We detail the challenges each of these methods addresses as well as the unique insights into aging that each has provided. We conclude with a discussion of potential future applications of these techniques as well as the importance of single-cell dynamics and quantitative biology approaches for understanding cell aging.

Effect of Opioid Limiting Legislation on Postoperative Opioid Utilization in Patients Treated for Ankle Fractures.

BACKGROUND: The state of Ohio implemented legislation in August of 2017 limiting the quantity of opioids a provider could prescribe. The purpose of this study was to identify if implementation of legislation affected opioid and nonopioid utilization in patients operatively treated for ankle fractures in the initial 90-day postoperative period after controlling for injury severity and preoperative narcotic usage. METHODS: A retrospective review of 144 patients treated for isolated ankle fractures in a pre-law group (January 2017-July 2017; n = 73) and post-law group (January 2018-July 2018; n = 71) was completed using electronic medical records and a legal prescriber database. Total number of opioid prescriptions, pills, milligrams of morphine equivalents (MMEs), and nonopioid prescriptions were recorded. Multiple regression analysis was run to identify predictors of opioid prescribing after controlling for law group, demographic, preoperative narcotic use, and injury severity characteristics. RESULTS: Mean MME prescribed per patient significantly decreased from 817.2 MME pre-law to 380.9 post-law (P < .01). Mean number of opioid pills prescribed per patient decreased from 99.1 in the pre-law group and 55.3 in the post law group (P < .001), respectively. Multiple linear regression analysis to predict the mean number of opioid pills prescribed was statistically significant (R 2 = 0.33; P < .001), with law group adding significantly to the prediction (P < .001). The multiple linear regression analysis to predict MME per patient was found to be statistically significant (R 2 = 0.31; P < .001), with the law group contributing significantly (P < .001). CONCLUSION: The Ohio prescriber law successfully contributed to the decreased number of opioid pills and MME prescribed in the initial 90-day postoperative period after controlling for injury severity and preoperative narcotic usage. Policies on opioid prescriptions may serve as an important public health tool in the fight against the opioid epidemic. LEVEL OF EVIDENCE: Level III, retrospective comparative series.

Divergent Aging of Isogenic Yeast Cells Revealed through Single-Cell Phenotypic Dynamics.

Although genetic mutations that alter organisms' average lifespans have been identified in aging research, our understanding of the dynamic changes during aging remains limited. Here, we integrate single-cell imaging, microfluidics, and computational modeling to investigate phenotypic divergence and cellular heterogeneity during replicative aging of single S. cerevisiae cells. Specifically, we find that isogenic cells diverge early in life toward one of two aging paths, which are characterized by distinct age-associated phenotypes. We captured the dynamics of single cells along the paths with a stochastic discrete-state model, which accurately predicts both the measured heterogeneity and the lifespan of cells on each path within a cell population. Our analysis suggests that genetic and environmental factors influence both a cell's choice of paths and the kinetics of paths themselves. Given that these factors are highly conserved throughout eukaryotes, divergent aging might represent a general scheme in cellular aging of other organisms.

Patterns in Opioid Prescription for Patients Operatively Treated for Ankle Fractures Following Implementation of 2017 Ohio Opioid Prescriber Law.

BACKGROUND: The purpose of this study was to report patterns of opioid prescription for patients treated operatively for ankle fractures after implementation of the 2017 Ohio Opioid Prescriber Law in comparison to the previous year. METHODS: A total of 144 patients operatively treated for isolated ankle fractures during two 6-month periods, January 2017 to July 2017 (pre-law) and January 2018 to July 2018 (post-law), were retrospectively identified. Preoperative and postoperative patient narcotic use was reviewed using a legal prescriber database. Total number of prescriptions, quantity of pills, and morphine milligram equivalents (MMEs) per patient prescribed during the 90-day postoperative period were compared between those treated before and those treated after implementation of the Ohio prescriber law. RESULTS: The average number of opioid prescriptions prescribed per patient in the 90-day postoperative period was 2.3 in the pre-law group and 2.1 in the post-law group (P = .625). The average MMEs prescribed per patient dropped from 942.4 MME pre-law to 700.5 MME post-law (P = .295). Differences in the average number of pills per prescription pre- and post-law (49.7 vs 36.2) and average MME per prescription (382.1 mg vs 275.2 mg) were statistically significant (P < .001 and P = .016, respectively). CONCLUSION: Following the implementation of the 2017 Ohio Opioid Prescriber Law, there was a downward trend in the number of pills per prescription and MMEs per prescription in patients operatively treated for isolated ankle fractures. The presence of a downward trend in the quantity of opioids prescribed in this patient cohort suggests the effectiveness of the state law. LEVEL OF EVIDENCE: Level III, comparative study.