Definitive internal fracture fixation followed by staged free flap coverage ("fix followed by flap" protocol) for open Gustilo type IIIB fractures.

BACKGROUND: Although the concept of the "fix and flap" approach, in which definitive fracture fixation and flap coverage are completed in a single procedure at the earliest opportunity may seem ideal for the treatment of Gustilo type IIIB open fractures, the individual circumstances of patients, such as polytrauma or multiple fracture cases may not allow for the immediate fracture fixation and flap coverage ("fix and flap" approach). In our hospital, patients with Gustilo type IIIB open fractures are treated with definitive internal fixation of the fracture followed by staged flap coverage ("fix followed by flap" protocol) when the "fix and flap" approach was not feasible due to the patient's condition or difficulty in coordinating surgery schedules. The "fix followed by flap" protocol provides benefits in terms of flexibility in adjusting the surgical timetable, simplifying the planning of flap coverage following fracture fixation, and minimizing individual surgical invasion. METHODS: We reviewed 10 cases of severe open fractures treated with the "fix followed by flap" protocol and evaluated their outcomes. All surgical procedures, including wound debridement, fracture fixation, and flap coverage, were performed by orthoplastic surgeons specializing in both fracture surgery and microsurgery including soft tissue reconstruction. RESULTS: All free flaps survived, and no partial necrosis was observed. None of the patients developed postoperative deep infection up to the last follow-up. Fracture union was achieved in all patients with or without autologous bone grafts. The median time for union was 9.4 months (range, 4-12 months). CONCLUSIONS: This study presents favorable outcomes of treatment for Gustilo type IIIB open fractures with fracture fixation followed by staged flap coverage ("fix followed by flap" protocol). Despite a delay in flap coverage, the consistency of treatment provided by orthoplastic surgeons may have contributed to the favorable outcomes in this study.

Bone and soft tissue reconstruction with tendon balance in severe foot trauma.

In severe foot trauma, it is difficult to determine the level of amputation when the crush injury is severe. We report a case of amputation near Lisfranc that achieved forefoot amputation-like results by using bone and soft tissue reconstruction while considering tendon balance. The patient was a 40-year-old male. The patient's left leg was caught in a garbage truck and sustained a crush injury. The second to fifth metatarsals were amputated at the diaphysis, and a high degree of instability of the Lisfranc joint was observed. A high degree of contamination was detected in the patient's wound, and the second to fifth toes were amputated at the Lisfranc joint during the initial treatment. The ends of the tibialis anterior, tibialis posterior and peroneus longus were preserved. On day 5 in the hospital, Lisfranc joint fixation of the hallux, amputation of the first metatarsal and reconstruction of the peroneus brevis were performed. On day 13, extensor hallucis longus tendon transfer and free anterolateral thigh flap were performed. On day 80, the patient was able to walk in regular shoes or non-orthopedic shoes. One year after reconstructive surgery, the patient had an average SAFE-Q score of 86.2 and mild varus deformity of the foot remained. In cases of severe foot trauma, the aim of reconstruction should be partial forefoot amputation whenever possible.

Cross-Limb Vascular Shunting for Traumatic Popliteal Artery Injury.

BACKGROUND: Popliteal artery injury (PAI) is a challenging trauma that requires prompt and accurate treatment since the probability of lower-limb amputation increases with the ischemic time. Intravascular shunting and cross-limb vascular shunting (CLS) are used as temporary vascular shunting (TVS) methods to shorten the ischemic time for limb vascular injury. CLS involves sending blood from an artery in a healthy body part to a peripheral vessel in an injured part to immediately resume blood flow to the injured limb. For closed injuries including PAI, CLS may be performed without exploring and identifying the arterial stumps and it enables early reperfusion to the ischemic limb. We report the case series of traumatic PAI treated using CLS and verify the usefulness of CLS. METHODS: All patients with traumatic PAI treated with CLS at our institution between August 2013 and December 2021 were included. Demographic and clinical patient characteristics were extracted from the medical records. Comorbid injuries, severity of acute limb ischemia based on the Rutherford grading scale, time from injury to reperfusion by CLS, time from injury to completion of artery, and the use of fasciotomy were investigated. As outcomes, we investigated the presence or absence of lower extremity amputation during the course of treatment. RESULTS: We used CLS as treatment for 5 cases with traumatic PAI. Based on the Rutherford grading scale for acute limb ischemia, there were one limb with grade 2B and 4 with grade 3. Amputation of the lower extremities was avoided except for 1 extremity in which arterial reconstruction was not achieved due to unexplained cardiac arrest during surgery. CONCLUSIONS: CLS enables early reperfusion of the injured limb and is effective as a TVS method for traumatic PAI with severe ischemia or soft tissue damage.

Management of bone loss in acute severe open tibial fractures: a retrospective study of twenty nine cases-a treatment strategy with bone length preservation.

PURPOSE: The present study investigated the outcomes of bone loss associated with acute open tibial fractures classified as Gustilo-Anderson classification grade III B (GIIIB) using a bone length preservation strategy. METHODS: Among acute GIIIB open tibial fractures, 29 limbs of 29 patients requiring bone loss treatment were included. The reconstruction methods for bone loss were selected among the Masquelet technique (MT), bone transport (BT), acute shortening followed by gradual lengthening (ASGL), and free vascularized fibula graft (FVFG). Primary outcome measures were the rate of bone union and time to bone union. RESULTS: The median radiographic apparent bone gap (RABG) was 46.75 mm. Bone loss was treated with ASGL only in two patients in whom it was not possible to cover large soft tissue defects by a single free latissimus dorsi (LD) myocutaneous flap (with the serratus anterior (SA) muscle). The other 27 patients underwent soft tissue reconstruction and bone loss treatment with the preservation of bone length, including the MT for 23, BT for six, and FVFG for one. The bone union rate was 75.9%, and the median time to bone union was six months. Salvage surgeries were performed on all seven patients with nonunion; all of whom eventually achieved bony union. CONCLUSION: Bone loss associated with acute GIIIB open tibial fractures were treated with "bone length preservation" if the size of the soft tissue defect was less than the size that was covered by a single LD myocutaneous flap (with the SA muscle).

Risk factors for over-telescoping in reverse oblique intertrochanteric fractures.

PURPOSE: Postoperative over-telescoping (OT) with lag screws is often observed in reverse oblique intertrochanteric fractures. This study aimed to clarify the risk factors of OT in patients with reverse oblique intertrochanteric fractures. METHODS: Electronic medical records of patients diagnosed with reverse oblique intertrochanteric fractures using plain radiography who underwent operative fixation with an intramedullary nail between August 2013 and December 2019 were reviewed. Patients were classified into two groups according to the Futamura classification: lateral wall pattern (LW) and reverse oblique pattern (RO). The incidence of OT in the LW and RO groups was compared. Also, we compared the incidence of OT for each reduction type in the LW group. RESULTS: Twenty patients had LW, and nine had RO. OT was observed in eight fractures (42.1%) in the LW group but not in the RO group. The incidence of OT was significantly higher in the LW group than in the RO group (P = 0.0261). Among the 19 fractures with LW, OT was observed in 7 of 10 and 1 of 9 fractures with postoperative reduction in the intramedullary and extramedullary or anatomical types, respectively. In the LW group, the incidence of OT was significantly higher in fractures with postoperative reduction in the intramedullary type than in those of the extramedullary or anatomical type (P = 0.0198). CONCLUSION: Our study showed that the incidence of OT was significantly higher in LW than in RO and that postoperative reduction in the intramedullary type in LW was a risk factor for OT.

Mechanism of action of non-camptothecin inhibitor Genz-644282 in topoisomerase I inhibition.

Topoisomerase I (TOP1) controls the topological state of DNA during DNA replication, and its dysfunction due to treatment with an inhibitor, such as camptothecin (CPT), causes replication arrest and cell death. Although CPT has excellent cytotoxicity, it has the disadvantage of instability under physiological conditions. Therefore, new types of TOP1 inhibitor have attracted particular attention. Here, we characterised the effect of a non-camptothecin inhibitor, Genz-644282 (Genz). First, we found that treatment with Genz showed cytotoxicity by introducing double-strand breaks (DSBs), which was suppressed by co-treatment with aphidicolin. Genz-induced DSB formation required the functions of TOP1. Next, we explored the advantages of Genz over CPT and found it was effective against CPT-resistant TOP1 carrying either N722S or N722A mutation. The effect of Genz was also confirmed at the cellular level using a CPT-resistant cell line carrying N722S mutation in the TOP1 gene. Moreover, we found arginine residue 364 plays a crucial role for the binding of Genz. Because tyrosine residue 723 is the active centre for DNA cleavage and re-ligation by TOP1, asparagine residue 722 plays crucial roles in the accessibility of the drug. Here, we discuss the mechanism of action of Genz on TOP1 inhibition.

One incision-two windows approach for fixation of multifragmentary coronoid process fracture of the ulna: A case report.

Since the range of access of each surgical approach around the elbow has limitations, it is difficult to treat all types of fractures using only one approach. In the case reported herein, anterior and medial fragments of the comminuted ulnar coronoid process fracture were treated by preparing two access routes through one skin incision and effectively performing the buttress plating of each fragment. The subject was a 27-year-old female who sustained a fracture of the coronoid process of the right ulna by falling during snowboarding. Computed tomography showed the concurrence of a type 2 subtype III and type 3 subtype I ulnar coronoid process fracture according to the O'Driscoll classification. The coronoid process was split into 3 parts: a fragment consisting of the anteromedial facet and upper half of the sublime tubercle (fragment 1), a central fragment including the tip (fragment 2), and a fragment extending from the radial side of the tip to the base of the coronoid process (fragment 3). A 12-cm-long skin incision was made on the anteromedial side of the elbow joint. The region of the anteromedial facet and sublime tubercle was reached by passage between the palmaris longus/flexor digitorum superficialis and humeral head of flexor carpi ulnaris using the over-the-top approach. Fragment 1 was fixed with a buttress plate. Using the anterior approach, the brachialis was then longitudinally split through by passage between the biceps and neurovascular bundle, fragments 2 and 3 were fixed together with a buttress plate. The "one incision-two windows" approach, which provides two approaches (the over-the-top window and the anterior window) by a single skin incision, was implemented for a multifragmentary ulnar coronoid process fracture. This approach is considered to offer access from the front to each of the anterior and medial fragments and permits appropriate buttress plate fixation.

Pull-in suture: a novel reconstruction technique for tendon avulsion injury at the musculotendinous junction associated with forearm open fracture.

We present three cases of strong one-staged tendon reconstruction for musculotendinous junction avulsion tendon injuries, and called it a 'pull-in suture'. The clinical outcomes of this method are comparable to those of tendon transfer; it is an effective reconstruction method that should be considered as an initial treatment procedure.

Prostaglandin EP4 Selective Agonist AKDS001 Enhances New Bone Formation by Minimodeling in a Rat Heterotopic Xenograft Model of Human Bone.

To enhance bone regeneration, the use of bone morphogenetic protein (BMP)-2 is an attractive option. Unfortunately, the dose-dependent side effects prevent its widespread use. Therefore, a novel osteogenic agent using a different mechanism of action than BMP-2 is highly desirable. Previous reports demonstrated that prostaglandin E2 receptor 4 (EP4) agonists have potent osteogenic effects on non-human cells and are one of the potential alternatives for BMP-2. Here, we investigated the effects of an EP4 agonist (AKDS001) on human cells with a rat heterotopic xenograft model of human bone. Bone formation in the xenograft model was significantly enhanced by AKDS001 treatment. Histomorphometric analysis showed that the mode of bone formation by AKDS001 was minimodeling rather than remodeling. In cultured human mesenchymal stem cells, AKDS001 enhanced osteogenic differentiation and mineralization via the cAMP/PKA pathway. In cultured human preosteoclasts, AKDS001 suppressed bone resorption by inhibiting differentiation into mature osteoclasts. Thus, we conclude that AKDS001 can enhance bone formation in grafted autogenous bone by minimodeling while maintaining the volume of grafted bone. The combined use of an EP4 agonist and autogenous bone grafting may be a novel treatment option to enhance bone regeneration. However, we should be careful in interpreting the results because male xenografts were implanted in male rats in the present study. It remains to be seen whether females can benefit from the positive effects of AKDS001 MS by using female xenografts implanted in female rats in clinically relevant animal models.

In vitro hemocompatibility investigation for the development of low-flow centrifugal blood pumps with less platelet clogging.

Low-flow blood pumps rated under 1 L/min are emerging for new medical applications, such as hemofiltration in acute use. In those pumps, platelet adhesion and aggregation have to be carefully considered because of clogging risk in the filter part. To find an acceptable hemocompatibility that can be applied to low-flow centrifugal blood pump design, the platelet aggregation index, clogging on a micromesh filter, and the hemolysis index were investigated using a low-flow blood pump designed for hemofiltration use. We conducted circulation testing in vitro using fresh porcine blood and two centrifugal pumps with different impeller inlet shapes. The Negative Log Platelet Aggregation Threshold Index (NL-PATI), which reflects the ability of residual platelets to aggregate, and flow rate were measured during reflux for 60 min, and the Normalized Index of Hemolysis (NIH (g/20 min)) was calculated. In addition, blood cell clogging after reflux was observed on the micromesh filter by SEM, and the adhesion rate was calculated. Our results showed that the platelet clogging on the micromesh filter occurred when the average NL-PATI was greater than 0.28 and the average NIH (g/20 min) was greater than 0.01. In contrast, platelet clogging on the micromesh was suppressed when NL-PATI was less than 0.17 and the NIH (g/20 min) was less than 0.003. These values might be used as acceptable hemocompatibility of low-flow centrifugal blood pumps with suppressed platelet clogging for hemofiltration pumps.

Quantitative investigation of platelet aggregation under high shear force for anti-platelet aggregation in vitro tests.

Blood pumps are often used for hemofiltration in patients with renal failure. To design effective centrifugal blood pumps for hemofiltration, it is important to suppress clogging caused by platelet aggregation. However, the optimal conditions for conducting anti-platelet aggregation tests in vitro have not yet been established. This study aimed to quantify the effect of the shear loading value and shear loading time on platelet aggregation and determine the optimal conditions for anti-platelet aggregation testing in vitro. To quantitatively evaluate platelet aggregation in terms of the negative logarithm-platelet aggregation threshold index (NL-PATI), which reflects the propensity of residual platelets to aggregate after shear loading, the following parameters were examined: blood collection method (collected from porcine vein using a syringe or collected from a slaughterhouse), type of anticoagulant (sodium citrate or heparin), shear rate, and shear time. The results showed that platelet aggregation in porcine blood increased under a high shear load applied at shear rates of approximately 20,000 s-1 or higher for 30 s. Platelet aggregation propensity was 2-3 times higher in heparin-anticoagulated blood than in sodium citrate-anticoagulated blood. Moreover, platelet aggregation was 1.5-2 times more in blood collected from the slaughterhouse than in syringe-collected blood. Testing with an integrated shear time of 30 s or less in relation to the total blood volume may be effective for conducting in vitro circulation experiments using hemofiltration blood pumps. The conditions established in this study may be useful for hemocompatibility testing of cardiovascular devices based on NL-PATI.

Improvement of hemolysis performance in a hydrodynamically levitated centrifugal blood pump by optimizing a shroud size.

We have developed a hydrodynamically levitated centrifugal blood pump. In the blood pump having hydrodynamic bearings, the narrow bearing gap has a potential for high hemolysis. The purpose of the this study is to improve hemolysis performance in a hydrodynamically levitated centrifugal blood pump by optimizing a shroud size. The impeller was levitated passively at the position where the thrust forces acting on the impeller were balanced. We focused on a size of a bottom shroud with a hydrodynamic bearing that could change the bottom hydrodynamic force to balance the thrust force at the wide bearing gap for reducing hemolysis. Five test models with various shroud size were compared: 989 mm2 (HH-10.5), 962 mm2 (HH-12), 932 mm2 (HH-13.5), 874 mm2 (HH-16), and 821 mm2 (HH-18). A numerical analysis was first performed to estimate the bearing gaps in the test model. The bearing gaps were then measured to validate the numerical analysis. Finally, an in vitro hemolysis test was performed. The numerical analysis revealed that the HH-13.5 model had the widest bearing gap of 129 µm. In the measurement test, the estimation error for the bearing gap was less than 10%. In the hemolysis test, the HH-13.5 model achieved the lowest hemolysis level among the five models. The present study demonstrated that the numerical analysis was found to be effective for determining the optimal should size, and the HH-13.5 model had the optimal shroud size in the developed hydrodynamically levitated centrifugal blood pump to reduce hemolysis.

Plasma skimming efficiency of human blood in the spiral groove bearing of a centrifugal blood pump.

This work investigates the plasma skimming effect in a spiral groove bearing within a hydrodynamically levitated centrifugal blood pump when working with human blood having a hematocrit value from 0 to 40%. The present study assessed the evaluation based on a method that clarified the limitations associated with such assessments. Human blood was circulated in a closed-loop circuit via a pump operating at 4000 rpm at a flow rate of 5 L/min. Red blood cells flowing through a ridge area of the bearing were directly observed using a high-speed microscope. The hematocrit value in the ridge area was calculated using the mean corpuscular volume, the bearing gap, the cross-sectional area of a red blood cell, and the occupancy of red blood cells. The latter value was obtained from photographic images by dividing the number of pixels showing red blood cells in the evaluation area by the total number of pixels in this area. The plasma skimming efficiency was calculated as the extent to which the hematocrit of the working blood was reduced in the ridge area. For the hematocrit in the circuit from 0 to 40%, the plasma skimming efficiency was approximately 90%, meaning that the hematocrit in the ridge area became 10% as compared to that in the circuit. For a hematocrit of 20% and over, red blood cells almost completely occupied the ridge. Thus, a valid assessment of plasma skimming was only possible when the hematocrit was less than 20%.

Real-time, non-invasive thrombus detection in an extracorporeal circuit using micro-optical thrombus sensors.

INTRODUCTION: Real-time, non-invasive monitoring of thrombus formation in extracorporeal circuits has yet to be achieved. To address the challenges of conventional optical thrombus detection methods requiring large devices that limit detection capacity, we developed a micro-optical thrombus sensor. METHODS: The proposed micro-optical thrombus sensor can detect the intensity of light scattered by blood at wavelengths of 660 and 855 nm. Two thrombus sensors were installed on in vitro circuit: one at the rotary blood pump and one at a flow channel. To evaluate the variation in the ratio of incident light intensity at each wavelength of the two sensors, Rfluct (for 660 nm) and Ifluct (for 855 nm) were defined. Using fresh porcine blood as a working fluid, we performed in vitro tests of haematocrit (Hct) and oxygen saturation (SaO2) variation and thrombus detection. Thrombus tests were terminated after Rfluct or Ifluct showed a larger change than the maximum range of those in the Hct and SaO2 variation test. RESULTS: In all three thrombus detection tests, Ifluct showed a larger change than the maximum range of those in the Hct and SaO2 variation test. After the tests, thrombus formation was confirmed in the pump, and there was no thrombus in the flow channel. The results indicate that Ifluct is an effective parameter for identifying the presence of a thrombus. CONCLUSION: Thrombus detection in an extracorporeal circuit using the developed micro-optical sensors was successfully demonstrated in an in vitro test.

Suitable hemolysis index for low-flow rotary blood pumps.

A suitable index is needed for hemolysis tests that use low-flow pumps, such as pediatric blood pumps or blood purification pumps. To create such an index, the present study investigates the change of plasma-free hemoglobin in the pump circuit with time and the change of the hemolysis rate with flow rate and impeller rotational speed. The results show that the hemolysis rate or the increase rate of the total free hemoglobin are suitable measures for hemolysis evaluation for low-flow pumps.

Bilateral Internal Carotid and Left Vertebral Artery Dissection after Blunt Trauma: A Case Report and Literature Review.

Multi-vessel cervical arterial injury after blunt trauma is rare, and its pathophysiology is unclear. Although blunt cerebrovascular injury is a common cause of cerebral ischemia, its management is still controversial. We describe a 23-year-old man in previously good health who developed three-vessel cervical arterial dissections due to blunt trauma. He was admitted to our emergency and critical care center after a motor vehicle crash. Computed tomography showed a thin, acute subdural hematoma in the right hemisphere and fractures of the odontoid process (Anderson type III), pelvis, and extremities. He was treated conservatively, and about 1 month later, he developed bleariness. Computed tomography angiography showed bilateral internal carotid and left vertebral artery dissection. Aspirin therapy was started immediately, and then clopidogrel was added to the regimen. Two weeks later, magnetic resonance angiography (MRA) showed improved blood flow of the vessels. Only aspirin therapy was continued. About 3 months after discharge, MRA demonstrated further improvement of the blood flow of both internal carotid arteries, but the dissection flap on the right side remained. Therefore, we extended the duration of antiplatelet therapy. On the basis of our experience with this case, we think that antithrombotic therapy is crucial for the management of multi-vessel cervical arterial injury, and agents should be used properly according to the injury grade and phase; however, further study is needed to confirm this recommendation.

Epidemiological State, Predictors of Early Mortality, and Predictive Models for Traumatic Spinal Cord Injury: A Multicenter Nationwide Cohort Study.

STUDY DESIGN: Multi-center, retrospective cohort study. OBJECTIVE: To determine the epidemiology, identify predictors of early mortality, and develop predictive models for traumatic spinal cord injury (SCI). SUMMARY OF BACKGROUND DATA: Despite improved initial care and management strategies, traumatic SCI remains a devastating event. Knowledge of the epidemiological state and predictive factors for mortality is important for developing strategies and counseling; however, they have not been adequately investigated, and predictive modeling regarding outcomes remains an underused modality for patients with traumatic SCI. METHODS: Using a nationwide trauma registry-the Japan Trauma Data Bank-we identified adult (≥18 yrs) patients with SCI between 2004 and 2015. The endpoint was in-hospital mortality. Characteristics of each patient were described. Multivariate logistic regression analyses were performed to identify factors significantly associated with in-hospital mortality and develop a predictive model. RESULTS: In total, 236,698 patients were registered in the database. Of the 215,835 adult patients, 8069 (3.7%) had SCI. The majority had SCI at the cervical level with falls at ground level being the primary etiology. Over the study period, median age, the proportion of cervical SCI, and the etiology of falls at ground level increased. The mortality rate was 5.6%. The following eight factors, age, sex, Glasgow Coma Scale on arrival (GCS), hypotension on arrival, bradycardia on arrival, severe head injury, Injury Severity Score (ISS), and neurological severity of SCI, were independently associated with mortality. A predictive model consisting of these variables predicted mortality with area under the receiver operating characteristic curve of 0.88 (95% confidence interval [CI], 0.86-0.90). CONCLUSION: Over the 12-year period, patient characteristics, etiology, and post-SCI outcomes significantly changed. We identified eight prognostic factors of early mortality. A predictive model including these factors showed excellent performance and may improve treatment strategies, healthcare resource allocation, and counseling. LEVEL OF EVIDENCE: 3.

Multi-scale instrumental analyses of plasticized polyhydroxyalkanoates (PHA) blended with polycaprolactone (PCL) and the effects of crosslinkers and graft copolymers.

Details of the mechanism underlying the tensile properties of plasticized polyhydroxyalkanoates (PHA) including poly(butylene succinate) (PBS) were investigated by blending with poly(ε-caprolactone) (PCL) as well as the addition of compatibilizers. Multi-scale instrumental analyses employed micro-focus X-ray CT to provide micro-scale morphology information on the order of ten microns while solid-state NMR spectral and relaxation time analyses contributed knowledge of the environment and molecular mobility of each constituent at the molecular to nano-scale. The blend of plasticized PHA with 50% PCL adopted a sea-island morphology to improve elongation at break in a quasi-static tensile test, which was dominated by the tensile properties of the added PCL. However, impact tensile properties were less improved by PCL addition, because its molecular mobility was suppressed by blending. Meanwhile, peroxy crosslinkers changed the sea-island morphology to homogenous in X-ray CT observations. Although the homogenous morphology sharply lowered the elongation at break in a quasi-static tensile test, the homogenous morphology improved impact tensile properties. Furthermore, graft polymers having acrylonitrile-styrene side-chains did not change the sea-island morphology but increased the molecular mobility of PBS in the plasticized PHA. This weak interaction between the plasticized PHA and PCL improved tensile properties in both quasi-static and impact tensile tests.

Discovery of a novel 2-(1H-pyrazolo[3,4-b]pyridin-1-yl)thiazole derivative as an EP1 receptor antagonist and in vivo studies in a bone fracture model.

We describe a medicinal chemistry approach to the discovery of a novel EP1 antagonist exhibiting high potency and good pharmacokinetics. Our starting point is 1, an EP1 receptor antagonist that exhibits pharmacological efficacy in cystometry models following intravenous administration. Despite its good potency in vitro, the high lipophilicity of 1 is a concern in long-term in vivo studies. Further medicinal chemistry efforts identified 4 as an improved lead compound with good in vitro ADME profile applicable to long term in vivo studies. A rat fracture study was conducted with 4 for 4 weeks to validate its utility in bone fracture healing. The results suggest that this EP1 receptor antagonist stimulates callus formation and thus 4 has potential for enhancing fracture healing.