Exogenous PTH 1-34 Attenuates Impaired Fracture Healing in Endogenous PTH Deficiency Mice via Activating Indian Hedgehog Signaling Pathway and Accelerating Endochondral Ossification

Fracture healing is a complicated, long-term, and multistage repair process. Intermittent administration of parathyroid hormone (PTH) has been proven effective on intramembranous and endochondral bone formation during the fracture healing process, however, the mechanism is unclear. In this study, we investigated the role of exogenous PTH and endogenous PTH deficiency in bone fracture healing and explored the mechanism by using PTH knockout (PTH-/-) mice and ATDC5 cells. In a mouse femur fracture model, endogenous PTH deficiency could delay endochondral ossification whereas exogenous PTH promotes accumulation of endochondral bone, accelerates cartilaginous callus conversion to bony callus, enhances maturity of bony callus, and attenuates impaired fracture healing resulting from endogenous PTH deficiency. In fracture callus tissue, endogenous PTH deficiency could inhibit chondrocyte proliferation and differentiation whereas exogenous PTH could activate the IHH signaling pathway to accelerate endochondral ossification and rescue impaired fracture healing resulting from endogenous PTH deficiency. In vitro, exogenous PTH promotes cell proliferation by activating IHH signaling pathway on ATDC5 cells. In mechanistic studies, by using ChIP and luciferase reporter assays, we showed that PTH could phosphorylate CREB, and subsequently bind to the promoter of IHH, causing the activation of IHH gene expression. Therefore, results from this study support the concept that exogenous PTH 1-34 attenuates impaired fracture healing in endogenous PTH deficiency mice via activating the IHH pathway and accelerating endochondral ossification. Hence, the investigation of the mechanism underlying the effects of PTH treatment on fracture repair might guide the exploration of effective therapeutic targets for fracture.

Systemic Administration of Recombinant Irisin Accelerates Fracture Healing in Mice

To date, pharmacological strategies designed to accelerate bone fracture healing are lacking. We subjected 8-week-old C57BL/6 male mice to closed, transverse, mid-diaphyseal tibial fractures and treated them with intraperitoneal injection of a vehicle or r-irisin (100 µg/kg/weekly) immediately following fracture for 10 days or 28 days. Histological analysis of the cartilaginous callus at 10 days showed a threefold increase in Collagen Type X (p = 0.0012) and a reduced content of proteoglycans (40%; p = 0.0018). Osteoclast count within the callus showed a 2.4-fold increase compared with untreated mice (p = 0.026), indicating a more advanced stage of endochondral ossification of the callus during the early stage of fracture repair. Further evidence that irisin induced the transition of cartilage callus into bony callus was provided by a twofold reduction in the expression of SOX9 (p = 0.0058) and a 2.2-fold increase in RUNX2 (p = 0.0137). Twenty-eight days post-fracture, microCT analyses showed that total callus volume and bone volume were increased by 68% (p = 0.0003) and 67% (p = 0.0093), respectively, and bone mineral content was 74% higher (p = 0.0012) in irisin-treated mice than in controls. Our findings suggest that irisin promotes bone formation in the bony callus and accelerates the fracture repair process, suggesting a possible use as a novel pharmacologic modulator of fracture healing.

Bony callus stiffness indirectly evaluated by the axial load-share ratio in vivo as a guide to removing a monolateral external fixator safely

Purpose As the monolateral external fixator is increasingly used in trauma-control and definitive management for high-energy long bone fractures, timing the fixator removal remains a challenge for surgeons. The purpose of this study was to determine the feasibility and effectiveness of the bony callus stiffness indirectly evaluated by the axial load-share ratio in vivo as a guide to removing a monolateral external fixator safely. Methods A total of 131 patients with tibial shaft fractures treated by the monolateral external fixator in our institution were collected from January 2013 to July 2019. In group I, the fixators were removed based on the clinical and radiological assessment only by the treating surgeon. As for group II, the axial load-share (LS) ratio test was accomplished by another medical team without the knowledge of the clinical results. The external fixator was removed when the mechanical test outcome (LS ratio < 10%) was consistent with the conclusion drawn from the clinical and radiological assessment (bone union achieved) by the treating surgeon. Results There was no statistical significance in demographic data between the two groups (P > 0.05). In group I, four patients suffered refracture (the refracture rate was 7.7%) after fixator removal and were successfully treated by an intramedullary nail. In group II, 71 patients underwent fixator removal after the first mechanical test, and another eight patients terminated the external fixation after the second test. None of the 79 patients in group II suffered refracture (the refracture rate was 0%). There was statistical significance in the refracture rate between the two groups (P < 0.05). Conclusion The bony callus stiffness indirectly evaluated by the axial load-share ratio in vivo using the additional circular frame components is an effectively quantitative indicator to complement the clinical assessment of fracture healing in a monolateral external fixation treatment. Removal of the monolateral external fixator is safe when the axial load-share ratio dropped below 10%.

The First Healed Bite Mark and Embedded Tooth in the Snout of a Middle Permian Gorgonopsian (Synapsida: Therapsida)

Despite their significance for paleobiological interpretations, bite marks have been rarely reported in non-mammalian therapsids (NMT). Here we describe, for the first time, the occurrence of a tooth embedded in the snout of a gorgonopsian. The tooth is surrounded by a bony callus, which demonstrates that the animal was still alive after the attack and healed. The identity of the attacker is unknown. Two hypotheses are discussed to account for this healed bite: failed predation (most likely by a biarmosuchian, therocephalian, or another gorgonopsian) and intraspecific social biting. Though predation cannot be ruled out, it has been hypothesized that gorgonopsians used their saber-like teeth for social signaling, which suggests that social biting may be the most likely scenario. The practice of social biting has long been hypothesized in NMT, but this is the first fossilized evidence of the behavior to be described.

Preliminary image findings of lower limb stress fractures to aid ultrasonographic diagnoses: A systematic review and narrative synthesis

Introduction This systematic review investigates which image appearances are most common when diagnosing lower limb stress fractures using ultrasound imaging, with the aim of outlining an image critique guideline for operators to support confident diagnoses. Method A comprehensive literature search of medical databases and handsearching was undertaken to identify relevant studies. All studies were critically examined for quality using the CASP critical appraisal tool. Results from eight studies were combined and interpreted using a narrative synthesis. Findings A clear outline of common stress fracture appearances using ultrasound were identified in a combined total of 119 participants. Each finding was ranked according to its popularity. Periosteal thickening (78/119) and cortical disruption/irregularity (83/119) were noted in all eight studies. Hypervascularity of the periosteum visualised by colour Doppler imaging (66/119) was reported in six of the eight studies. Soft tissue hypervascularity (13/119), bony callus formation (5/119) and cortical break (22/119) were seen in three studies. Conclusions Based on the findings, we offer a guideline of the most significant preliminary image findings to be utilised by operators when examining athletes suspected of having lower limb stress fractures. The results show a gap in research for evaluating changes in appearance depending on the injury severity. Further research into distinguishing stress fractures from pathological involvement may in future reduce reliance on plain film radiography.

Arthrectomy for traumatic proximal interphalangeal arthritis in the lateral digit in a heifer

Instructive information on the treatment for chronic deep infections of digital joints in a 9-month-old Holstein heifer is described in this report. Preoperative ultrasonographic and radiographic examinations revealed soft tissue swelling and subchondral bone lysis at the distal part of the proximal phalanx in the lateral digit. Arthrectomy was performed under xylazine sedation to remove infectious articular cartilage tissues. Immature callus formation was observed via radiography at the surgical site by the 28th postoperative day. On the 48th postoperative day, callus fellinto disrepair on the radiographs along with aggravation of the locomotion score. After the application of a half-limb cast, the immature callus formed again by the 62nd postoperative day (11 months), and bony callus formation was observed by the 74th postoperative day. Thereafter, the heifer could walk well with marked improvement in the locomotion score. The withers height of the heifer at 13 months (136 cm) was within the range of that in control heifers of the same age on this farm (133 ± 3 cm); however, the body weight (BW) of this heifer (322 kg) was lower than the BW of controls (384 ± 26 kg). The BW gain from 11 to 13 months of age seemed to be higher in the present heifer (+76 kg) than in controls (+55±20 kg), suggesting that BW of the present heifer was returning to the original BW. Based on these observations, we suggested that arthrectomy was an effective treatment option for the present case of digital joint arthritis.

Impact of the Sensory and Sympathetic Nervous System on Fracture Healing in Ovariectomized Mice

The peripheral nervous system modulates bone repair under physiological and pathophysiological conditions. Previously, we reported an essential role for sensory neuropeptide substance P (SP) and sympathetic nerve fibers (SNF) for proper fracture healing and bone structure in a murine tibial fracture model. A similar distortion of bone microarchitecture has been described for mice lacking the sensory neuropeptide α-calcitonin gene-related peptide (α-CGRP). Here, we hypothesize that loss of SP, α-CGRP, and SNF modulates inflammatory and pain-related processes and also affects bone regeneration during fracture healing under postmenopausal conditions. Intramedullary fixed femoral fractures were set to 28 days after bilateral ovariectomy (OVX) in female wild type (WT), SP-, α-CGRP-deficient, and sympathectomized (SYX) mice. Locomotion, paw withdrawal threshold, fracture callus maturation and numbers of TRAP-, CD4-, CD8-, F4/80-, iNos-, and Arg1-positive cells within the callus were analyzed. Nightly locomotion was reduced in unfractured SP-deficient and SYX mice after fracture. Resistance to pressure was increased for the fractured leg in SP-deficient mice during the later stages of fracture healing, but was decreased in α-CGRP-deficient mice. Hypertrophic cartilage area was increased nine days after fracture in SP-deficient mice. Bony callus maturation was delayed in SYX mice during the later healing stages. In addition, the number of CD 4-positive cells was reduced after five days and the number of CD 8-positive cells was additionally reduced after 21 days in SYX mice. The number of Arg1-positive M2 macrophages was higher in α-CGRP-deficient mice five days after fracture. The alkaline phosphatase level was increased in SYX mice 16 days after fracture. Absence of α-CGRP appears to promote M2 macrophage polarization and reduces the pain threshold, but has no effect on callus tissue maturation. Absence of SP reduces locomotion, increases the pain-threshold, and accelerates hypertrophic callus tissue remodeling. Destruction of SNF reduces locomotion after fracture and influences bony callus tissue remodeling during the later stages of fracture repair, whereas pain-related processes are not affected.

Biomechanical Assessment for Healing Progression of Fractured Long Bones: Numerical Investigations of Bending Stiffness and Resonant Frequency

Though biomechanical methods have been proposed to assess the healing status of fractured bones for more than two decades, the effective stiffnesses obtained theoretically never match the experimentally measured results for the healing progression. This paper proposed a novel numerical model, the simulation results of which are able to reflect the experimental observations. The callus is divided into 10 regions with different material properties and the gap-narrowing process of the fracture is simulated with the variations of the material properties during the healing progression. The variations of the effective bending stiffness and resonant frequency with the healing progression have been obtained. The bending stiffness and resonant frequency squared of the fractured bone show a substantial creeping part during gradual narrowing of the fracture gap. They start to increase rapidly during bridging of the fracture gap by a bony callus. When the Young’s modulus in the callus region calcified last (i.e., Region 10) reaches 5% that of the reference intact bone, the bending stiffness and resonant frequency squared rise up to 90% those of the reference intact bone. After that they increase slowly and get flat; they become less sensitive to bone growth in the late healing stages. It is the first time that the simulation results demonstrate three distinct stages like in the experimentally measured results. These results imply that the variations of the bending stiffness and resonant frequency squared are not linearly correlated with the healing progression; however, their fast growing phase does indicate bony bridging of the callus.