Morphine-induced osteolysis and hypersensitivity is mediated through toll-like receptor-4 in a murine model of metastatic breast cancer.

ABSTRACT: The propensity for breast cancer to metastasize to bone is coupled to the most common complaint among breast cancer patients: bone pain. Classically, this type of pain is treated using escalating doses of opioids, which lack long-term efficacy due to analgesic tolerance, opioid-induced hypersensitivity, and have recently been linked to enhanced bone loss. To date, the molecular mechanisms underlying these adverse effects have not been fully explored. Using an immunocompetent murine model of metastatic breast cancer, we demonstrated that sustained morphine infusion induced a significant increase in osteolysis and hypersensitivity within the ipsilateral femur through the activation of toll-like receptor-4 (TLR4). Pharmacological blockade with TAK242 (resatorvid) as well as the use of a TLR4 genetic knockout ameliorated the chronic morphine-induced osteolysis and hypersensitivity. Genetic MOR knockout did not mitigate chronic morphine hypersensitivity or bone loss. In vitro studies using RAW264.7 murine macrophages precursor cells demonstrated morphine-enhanced osteoclastogenesis that was inhibited by the TLR4 antagonist. Together, these data indicate that morphine induces osteolysis and hypersensitivity that are mediated, in part, through a TLR4 receptor mechanism.

Robot and ultrasound assisted needle insertion to the transverse carpal ligament.

BACKGROUND: A potential alternative treatment to surgery for carpal tunnel syndrome is to inject enzymes into the transverse carpal ligament to decrease its stiffness and alleviate pressure off the median nerve. An accurate injection is needed for delivery to achieve the effects of tissue degradation. The purposes of this study were to 1) determine injection sites using 3D reconstructed anatomy, and 2) insert the needle to the middle of the transverse carpal ligament thickness in situ. METHODS: Six fresh-frozen cadaveric hands were used in this study. Five injection sites were determined in the sagittal plane along the center of the transverse carpal ligament thickness ulnar to the thenar muscle attachment using 3D ultrasonographic reconstruction. Each injection was delivered by rigidly fixing a 27-gauge needle to a six degrees of freedom robot arm programmed to insert the needle tip to the intended target. Ultrasound images were taken of the needle after insertion to measure accuracy and precision of the needle placement. FINDINGS: The needle tip was successfully delivered to the middle region of the transverse carpal ligament thickness and visualized using ultrasound imaging. The accuracy and precision of the needle insertion were 0.83 and 0.31 mm, respectively. INTERPRETATION: Methodology was established for robot-assisted needle insertion to the transverse carpal ligament using 3D ultrasonographic reconstructed anatomy. This methodology can be used in the future to deliver enzymatic injections to the transverse carpal ligament as a potential treatment for carpal tunnel syndrome.

A Large Segmental Mid-Diaphyseal Femoral Defect Sheep Model: Surgical Technique.

Background:There are numerous animal models available to study bone healing as well as test strategies to accelerate bone formation. Sheep are commonly used for evaluation of long bone fractures due to similar dimensions and weight bearing environments compared to patients. Large critical-size defects can be created in sheep to facilitate the study of implantable materials, osteogenic proteins, and stem cell treatments. Studies have been published using plates to stabilize large critical size defects in femoral, tibial, and metatarsal defects. External fixators have also been used to stabilize tibial defects in sheep.Methods: The purpose of the current paper is to detail the surgical technique for creation of a 42 mm mid-diaphyseal femoral defect stabilization with an intramedullary device in sheep. Additional surgical details are provided for dynamization, reverse dynamization, and device removal.Conclusion: The article provides multiple technical tips applicable to this and other ovine osteotomy models and concludes with a discussion comparing the use of each stabilization technique in clinically significant large critical-size bone defects.

Axonal loss in murine peripheral nerves following exposure to recombinant human bone morphogenetic protein-2 in an absorbable collagen sponge.

BACKGROUND: With the proven efficacy of recombinant human bone morphogenetic protein-2 (rhBMP-2) to treat open tibial fractures and promote spine fusion, there has been an increase in its off-label use. Recent studies have shown that BMPs play a role in nerve development and regeneration. Little is known about changes that result when rhBMP-2 is used in the vicinity of peripheral nerves. The purpose of this study is to characterize changes in peripheral nerves following exposure to rhBMP-2-soaked collagen sponges. METHODS: rhBMP-2 on an absorbable collagen sponge (ACS) was implanted directly on the sciatic nerves of Wistar rats. One and three weeks following surgery, the nerves were harvested and histological analysis was performed to evaluate inflammatory and structural changes. RESULTS: rhBMP-2-soaked collagen sponges induced ectopic bone formation in muscle tissue in all animals after three weeks, but did not cause bone formation within the nerve. Axonal swelling and splitting of the myelin sheath were observed in both experimental and control nerves and may be a result of surgical manipulation. The overall incidence of axonal loss was 15.8% in the rhBMP-2/ACS-exposed nerves and was 0% in control nerves (p < 0.05). CONCLUSIONS: rhBMP-2-soaked collagen sponges may adversely affect the axons of peripheral nerves by causing axonal dropout and loss of axons. Ectopic bone formation occurs within muscle tissues and not within the peripheral nerve. The axonal dropout may be a direct effect of rhBMP-2-soaked collagen sponges and not nerve compression as it was observed prior to ectopic bone formation.

Evaluation of the osteogenic performance of calcium phosphate-chitosan bone fillers.

There has been recent interest in utilizing calcium phosphates (CaPs) that set in situ for treating bone defects due to the limitations associated with morselized autografts and allografts. However, CaP cements have long setting times, poor mechanical properties, and poor osteoinductivity. This has prompted research toward finding a nonprotein-based compound, such as chitosan, to accelerate setting times and increase osteoinductivity. The purpose of this study was to compare bone growth rates during the early bone healing response achieved using conventionally prepared chitosan-CaP bone filler to an extensively purified chitosan-CaP compound. Both compounds set quickly and stimulated bone formation. Histomorphometry demonstrated a 290% increase in new bone formation when using the conventional chitosan-CaP bone filler and a 172% increase with the highly purified chitosan-CaP compound compared to the increase in bone formation seen with the unfilled control group. The results of this study indicate that a highly purified chitosan-CaP paste stimulated less bone formation than a conventionally prepared chitosan-CaP paste but both pastes have the potential to stimulate bone formation.

Load measurement accuracy from sensate scaffolds with and without a cartilage surface.

ABSTRACT The use of "sensate" scaffolds covered with tissue-engineered cartilage has emerged as a possible treatment option for focal articular cartilage defects. The ability to monitor joint loading provides several benefits that can be useful in both clinical and research situations. Previous studies have shown that these scaffolds can accurately monitor in vivo joint loading during various activities. However, the effect that an articular cartilage layer or soft tissue overgrowth has on scaffold sensitivity has not been tested. Eight scaffolds were tested with cartilage samples taken from four hounds. Three strain gauges were attached to each scaffold and a servo hydraulics system was used to test sensitivity while the scaffold was in contact with cartilage, metal, or silicone surfaces. Strain gauge sensitivity was calculated from load and strain measurements collected during testing. There was no significant difference between the mean strain gauge sensitivities when the scaffolds were in contact with the different surfaces: cartilage 30.9 +/- 16.2 muepsilon/N, metal 31.8 +/- 18.6 muepsilon/N, and silicone 30.6 +/- 12.3 muepsilon/N. These results indicate that "sensate" scaffolds can be calibrated and used to monitor load with the presence of an articular cartilage layer.

In vivo strain measurements from hardware and lamina during spine fusion.

Currently, spine fusion is determined using radiography and clinical evaluation. There are discrepancies between radiographic evidence and direct measurements of fusion, such as operative exploration and biomechanical or histological measurements. In order to facilitate the rapid return of patients to normal activities, a monitoring technique to accurately detect fusion in vivo and to prevent overload during the postoperative period would be useful. The objectives of this study were to develop an implantable monitoring system consisting of CPC-coated strain gauges and a radio transmitter to detect the onset of fusion and measure strain during postsurgical activities. A patient underwent anterior release and fusion, followed by posterior instrumentation and fusion with segmental spinal instrumentation. Four strain gauges were placed during surgery. One was attached to the left-side rod and one to each of the lamina at T9, T10, and T11. An externally powered implanted radio transmitter attached to the gauges was placed in a subcutaneous pouch. Strains were monitored weekly and tabulated during various activities for 7 months. Peak strains during twisting and bending were tabulated to detect the onset of fusion. Strains were also recorded during activities such as climbing off an examination table, rising from a chair, and climbing stairs. Strains collected from the left rod indicated that, immediately postoperatively, it was loaded at acceptable levels. The largest and most consistent strain changes measured from the lamina were recorded during twisting.