Biomimetic Metal-Organic Frameworks as Targeted Vehicles to Enhance Osteogenesis.

Although engineered nanoparticles loaded with specific growth factors are used to regulate differentiation of stem cells, the low loading efficiency and biocompatibility are still great challenges in tissue repair. A nature-inspired biomimetic delivery system with targeted functions is attractive for enhancing cell activity and controlling cell fate. Herein, a stem cell membrane (SCM)-wrapped dexamethasone (DEX)-loaded zeolitic imidazolate framework-8 (ZIF-8) is constructed, which integrates the synthetic nanomaterials with native plasma membrane, to achieve efficient DEX delivery and DEX-mediated bone repair. The DEX@ZIF-8-SCM enables high DEX loading capacity, modulates the sustained release, and facilitates the specific uptake of mesenchymal stem cells (MSCs), owing to the porous property of ZIF-8 and the innate targeting capability of SCM. The endocytosed DEX@ZIF-8-SCM shows high cytocompatibility and greatly enhances the osteogenic differentiation of MSCs. Furthermore, RNA-sequencing data reveal that the phosphoinositide 3-kinase (PI3K)-Akt signaling pathways are activated and dominantly involved in the accelerated osteogenesis. In the bone defect model, the administrated DEX@ZIF-8-SCM exerts excellent biocompatibility and effectively promotes bone regeneration. Overall, the SCM-derived biomimetic nanoplatform achieves targeted delivery, excellent biosafety, and enhanced osteogenic differentiation and bone repair, which provides a new and valid strategy for treating various tissue injuries.

Severe Leukemoid Reaction in a Case of Soft Tissue Sarcoma: A Case Report.

CASE: A 62-year-old woman presented with severe leukocytosis and systemic symptoms including fatigue, low appetite, and weight loss. After 4 months of evaluation by doctors with extensive laboratory tests and imaging studies, a soft tissue sarcoma in the left posterior distal thigh was revealed. Quickly after surgical excision of the soft tissue sarcoma, the leukocytosis and systemic symptoms were dramatically resolved. CONCLUSIONS: Leukemoid reaction can present as a paraneoplastic syndrome. Awareness of soft tissue sarcoma as a possible cause of leukemoid reaction may help improve the early diagnosis and subsequent early intervention in future cases.

Selective recovery of Ag(I) coordination anion from simulate nickel electrolyte using corn stalk based adsorbent modified by ammonia-thiosemicarbazide.

In nickel electrolyte, Ag(I) was present at trace level concentration (10-20 mg L(-1)) and existed in the form of AgCli(1-i) coordination anion, instead of Ag(+) positive ion usually in several sources. In the present study, TSC-NH3-OCS adsorbent based on natural corn stalk modified by ammonia (NH3)-thiosemicarbazide (TSC) was synthesized and characterized using some instrumental techniques. The TSC-NH3-OCS adsorbent could selectively adsorb Ag(I) as AgCl(i)(1-i) coordination anion from the Ag(I)-Cu(II)-Ni(II) simulate nickel electrolyte, especially in the case of the very high levels of Cu(II) and Ni(II), which significantly outperforms the commercial available resins. The adsorption mechanism was believed to be electrostatic interaction of the protonated bands of AgCl4(3-) with protonated thiol form of the thioamide units by FTIR and XPS analysis. The maximum adsorption capacity in the Ag(I) single and Ag(I)-Cu(II)-Ni(II) ternary system were obtained and calculated as 153.54 and 46.69 mg g(-1), respectively. The reasons that the maximum adsorption capacity of AgCl(i)(1-i) from the single and ternary system varied widely could be explained by adsorption kinetic and thermodynamic results. In addition, three successive sorption/desorption cycle runs from ternary system were performed which indicated that the TSC-NH3-OCS adsorbent has a good performance for recovery Ag(I) from simulate nickel electrolyte.

Pelvic crescent fractures: variations in injury mechanism and radiographic pattern.

Pelvic crescent fracture, also known as sacroiliac fracture-dislocation, is traditionally considered as a lateral compression injury and a vertically stable injury. Thirty consecutive cases were analyzed and it was found that 63% of cases were caused by lateral compression (LC), 27% by anteroposterior compression (APC), and 10% by vertical shear (VS). APC and VS injuries cause significant displacement of the anterior iliac fragment, but 21% of LC injury cases showed minimal displacement and were treated successfully with nonoperative treatment. Different injury mechanisms also produce different types of pelvic instability. More important, different injury mechanisms produce distinct radiographic fracture patterns regarding the obliquity of the fracture line and fracture surface. These differences in the fracture pattern will influence the decision of internal fixation options. Therefore, treatment of pelvic crescent fractures should be based on individual analysis of injury mechanism and radiographic fracture pattern.

A long femoral stem is not always required in hip arthroplasty for patients with proximal femur metastases.

BACKGROUND: During hip arthroplasties for treating proximal femur metastases, a long femoral stem frequently is used, presumably protecting the entire femur against progression of the existing lesions or development of new lesions. However, it is unclear whether a long stem is really required. QUESTIONS/PURPOSES: We therefore determined in patients with proximal femur metastases (1) the reoperation rate related to different stem lengths after hip arthroplasty, (2) the risk of tumor progression in the same femur (the progression of preexisting lesions and the development of new distal femur lesions), and (3) complications. METHODS: We retrospectively reviewed 203 patients (206 femurs) with proximal femur metastases treated with hip arthroplasty. These femurs were divided into three groups based on femoral stem length: short stem (SS), 12 to 14 cm; medium stem (MS), 20 to 24 cm; and long stem (LS), 25 to 35 cm. We reviewed reoperations, disease progression in the same femur, and complications. Minimum followup was 2 days (median, 487 days; range, 2-4853 days), with most patients followed to their death. RESULTS: Only three femurs were revised owing to tumor progression, with no difference among the SS, MS, and LS groups. Two SS prostheses were revised for nononcologic reasons. Tumor progression in the same femur was uncommon during the patient's survival, with 11 femurs showing progression of the proximal lesion and five femurs showing new distal lesions. The complication rate was higher in the LS group (28%) than the combined rate in the MS and SS groups (16%), especially acute cardiopulmonary complications (18% versus 7.5%). CONCLUSIONS: Reoperation after hip arthroplasty for proximal femur metastases is uncommon and not correlated with femoral stem length. Considering the high complication rate associated with a LS hip prosthesis, we do not believe its routine use is justified.

Unicompartmental knee arthroplasty: factors influencing the outcome.

Currently, the outcome and indications of unicompartmental knee arthroplasty (UKA) are still controversial. We retrospectively reviewed a consecutive case series of UKA done by a single surgeon between 2004 and 2007 including 178 knees (140 patients). There were 31 lateral UKA and 147 medial UKA. With a minimum follow-up of 24 months (average 54 months) in 159 knees (other 19 knees were lost to follow-up at 3 to 18 months after surgery), 6 knees (3.8%, all medial UKA) were converted to total knee arthroplasty in 17 to 66 months (average 33 months). We found that the outcome of UKA was not influenced by the patient's age, body mass index, or early degeneration in the patellofemoral joint (PFJ). Compared with other reports, there was a greater proportion of lateral UKA in our series (17.4%). Although lateral UKA showed a trend toward less complications and implant failure compared with medial UKA, WOMAC scores (Western Ontario and McMaster Universities Osteoarthritis Index) were similar between the two types of partial knee arthroplasty. Our results indicate that young age, obesity, and early degeneration in the PFJ may not be contraindications to UKA, and lateral UKA functions as well as, if not better than medial UKA. However, a long-term follow-up is required to confirm these findings.

Anti-inflammatory treatment increases angiogenesis during early fracture healing.

OBJECTIVES: Both inflammation and angiogenesis are crucial for normal fracture healing. The goal of this work was to determine how anti-inflammatory treatment affects angiogenesis during early stages of fracture repair. METHODS: Tibia fractures were created in adult mice and animals were treated with indomethacin (2 mg/kg/day), a non-steroidal anti-inflammatory drug, or PBS once a day beginning from 1 day before fracture and continuing to 6 days after fracture. Animals were killed at 7, 14, and 28 days after injury for histomorphometric analysis of fracture healing. A second group of animals were killed at 3 and 7 days after injury to measure tissue levels of VEGF and interleukin-1 beta (IL-1ß). A third group of animals were killed at 3 and 7 days after injury for stereology analysis of macrophage and neutrophil infiltration and tissue vascularization. RESULTS: Indomethacin significantly decreased bone and cartilage formation at 7 days after fracture compared to controls. Indomethacin decreased the tissue levels of IL-1ß at 3 days after fracture but did not affect the recruitment of macrophages or neutrophils to injured limbs. Indomethacin-treated fractures had similar length density and surface density of vasculature as the controls at 3 days after injury. At 7 days after fracture, vasculature in indomethacin-treated fractures exhibited higher length density and surface density than that in controls. By 28 days after injury, indomethacin-treated fractures still exhibited defects in fracture repair. CONCLUSIONS: Anti-inflammatory treatments using indomethacin impair bone and cartilage formation and increase tissue vascularization in the callus during early fracture healing.

Mechanical stability affects angiogenesis during early fracture healing.

OBJECTIVES: The goal of this study was to determine to what extent mechanical stability affects vascular repair during fracture healing. METHODS: Stabilized and nonstabilized tibia fractures were created in adult mice. Fracture tissues were collected at multiple time points during early fracture healing. Vasculature in fractured limbs was visualized by immunohistochemistry with an anti-PECAM-1 antibody on tissue sections and then quantified with stereology. Oxygen tension, vascular endothelial growth factor expression, and lactate accumulation at the fracture site were measured. Gene expression was compared between stabilized and nonstabilized fractures by microarray analysis. RESULTS: We found that new blood vessel formation was robust by 3 days after fracture. Quantitative analysis showed that nonstabilized fractures had higher length density and surface density than stabilized fractures at 3 days after injury, suggesting that nonstabilized fractures were more vascularized. Oximetry analysis did not detect a significant difference in oxygen tension at the fracture site between stabilized and nonstabilized fractures during the first 3 days after injury. Further microarray analysis was performed to determine the effects of mechanical stability on the expression of angiogenic factors. No significant difference in the expression of vascular endothelial growth factors and other angiogenic factors was detected between stabilized and nonstabilized fractures. CONCLUSIONS: Mechanical instability promotes angiogenesis during early fracture healing and further research is required to determine the underlying mechanisms.

Multiple roles for CCR2 during fracture healing.

Bone injury induces an inflammatory response that involves neutrophils, macrophages and other inflammatory cells. The recruitment of inflammatory cells to sites of injury occurs in response to specific signaling pathways. The CC chemokine receptor type 2 (CCR2) is crucial for recruiting macrophages, as well as regulating osteoclast function. In this study, we examined fracture healing in Ccr2-/- mice. We first demonstrated that the expression of Ccr2 transcripts and the filtration of macrophages into fracture calluses were most robust during the early phases of fracture healing. We then determined that the number of macrophages at the fracture site was significantly lower in Ccr2-/- mice compared with wild-type controls at 3 days after injury. As a result, impaired vascularization, decreased formation of callus, and delayed maturation of cartilage were observed at 7 days after injury in mutant mice. At day 14, Ccr2-/- mice had less bone in their calluses. At day 21, Ccr2-/- mice had larger calluses and more bone compared with wild-type mice, suggesting a delayed remodeling. In addition, we examined the effect of Ccr2 mutation on osteoclasts. We found that a lack of Ccr2 did not affect the number of osteoclasts within fracture calluses at 21 days after injury. However, Ccr2-/- osteoclasts exhibited a decreased ability to resorb bone compared with wild-type cells, which could contribute to the delayed remodeling of fracture calluses observed in Ccr2-/- mice. Collectively, these results indicate that a deficiency of Ccr2 reduces the infiltration of macrophages and impairs the function of osteoclasts, leading to delayed fracture healing.

Rejuvenation of the inflammatory system stimulates fracture repair in aged mice.

Age significantly reduces the regenerative capacity of the skeleton, but the underlying causes are unknown. Here, we tested whether the functional status of inflammatory cells contributes to delayed healing in aged animals. We created chimeric mice by bone marrow transplantation after lethal irradiation. In this model, chondrocytes and osteoblasts in the regenerate are derived exclusively from host cells while inflammatory cells are derived from the donor. Using this model, the inflammatory system of middle-aged mice (12 month old) was replaced by transplanted bone marrow from juvenile mice (4 weeks old), or age-matched controls. We found that the middle-aged mice receiving juvenile bone marrow had larger calluses and more bone formation during early stages and faster callus remodeling at late stages of fracture healing, indicating that inflammatory cells derived from the juvenile bone marrow accelerated bone repair in the middle-aged animals. In contrast, transplanting bone marrow from middle-aged mice to juvenile mice did not alter the process of fracture healing in juvenile mice. Thus, the roles of inflammatory cells in fracture healing may be age-related, suggesting the possibility of enhancing fracture healing in aged animals by manipulating the inflammatory system.

Recombinant human bone morphogenetic protein-7 enhances fracture healing in an ischemic environment.

Ischemia predisposes orthopedic trauma patients to delayed fracture healing or nonunion. The goal of this study was to test the ability of bone morphogenetic protein 7 (BMP7) to stimulate fracture repair in an ischemic environment. Ischemic fractures were generated in male adult mice by resecting the femoral artery prior to the creation of a nonstabilized tibia fracture. Recombinant human BMP7 (rhBMP7, 50 microg) was injected into the fracture site immediately after surgery. At 7 days after injury, more tissue vascularization was observed in rhBMP7 treated fractures. Histomorphometric analyses revealed that rhBMP7 induced more cartilage at day 7, more callus and bone at days 14 and 28, and more adipose tissue and fibrous tissue at days 7, 14, and 28 compared to controls (n=5/group/time). At day 28, all fractures treated with rhBMP7 (50 microg, n=5) healed, whereas only three of five control fractures exhibited slight bony bridging. In addition, we found that rhBMP7 (both 10 and 50 microg) significantly increased the amount of cartilage compared to controls in stabilized fractures, confirming its chondrogenic effect. Lastly, using bone marrow transplantation, we determined that no donor-derived osteocytes or chondrocytes were present in rhBMP7-treated fractures, suggesting rhBMP7 did not recruit mesenchymal stem cells from the bone marrow to the fracture site. In conclusion, our results indicate that rhBMP7 is a promising treatment for fractures with severely disrupted blood supply.

Titanium particles that have undergone phagocytosis by macrophages lose the ability to activate other macrophages.

Titanium particles derived from the wear of the orthopaedic implant surfaces can activate macrophages to secrete cytokines and stimulate osteoclastic bone resorption, causing osteolysis around orthopaedic implants. However, what happens to the titanium particles after being phagocytosed by macrophages is not known. We prepared titanium particles (as received, clean, and LPS-coated), and exposed them to macrophages in culture. Free particles were washed away after 24 h and the intracellular particles were kept in culture for additional 48 h until being harvested by lysing the cells. Particles that had been cell treated or noncell treated were examined by scanning electronic microscopy to analyze the shape, size, and concentration of the particles. The cell treated and noncell treated particles were exposed to macrophages in culture with a particle to cell ratio of 300:1. After 18 h, the levels of TNF-alpha in culture medium and the viability of the cells were examined. Clean particles did not stimulate TNF-alpha secretion by macrophages, while LPS-coated particles dramatically increased that response. Phagocytosis by macrophages did not change the shape and size of the particles, but depleted the ability of the particles to stimulate TNF-alpha secretion by macrophages. This indicates that macrophages are capable of rendering titanium particles inactive without degrading the particles, possibly by altering the surface chemistry of the particles.

Role of matrix metalloproteinase 13 in both endochondral and intramembranous ossification during skeletal regeneration.

Extracellular matrix (ECM) remodeling is important during bone development and repair. Because matrix metalloproteinase 13 (MMP13, collagenase-3) plays a role in long bone development, we have examined its role during adult skeletal repair. In this study we find that MMP13 is expressed by hypertrophic chondrocytes and osteoblasts in the fracture callus. We demonstrate that MMP13 is required for proper resorption of hypertrophic cartilage and for normal bone remodeling during non-stabilized fracture healing, which occurs via endochondral ossification. However, no difference in callus strength was detected in the absence of MMP13. Transplant of wild-type bone marrow, which reconstitutes cells only of the hematopoietic lineage, did not rescue the endochondral repair defect, indicating that impaired healing in Mmp13-/- mice is intrinsic to cartilage and bone. Mmp13-/- mice also exhibited altered bone remodeling during healing of stabilized fractures and cortical defects via intramembranous ossification. This indicates that the bone phenotype occurs independently from the cartilage phenotype. Taken together, our findings demonstrate that MMP13 is involved in normal remodeling of bone and cartilage during adult skeletal repair, and that MMP13 may act directly in the initial stages of ECM degradation in these tissues prior to invasion of blood vessels and osteoclasts.

Administration of pamidronate alters bone-titanium attachment in the presence of endotoxin-coated polyethylene particles.

Bisphosphonates are promising in the treatment of periprosthetic osteolysis induced by particulate wear debris. The in vivo effects of pamidronate with different doses and durations of administration on bone-titanium attachment in the presence of endotoxin-coated polyethylene particles were examined in a rat model in this study. Titanium pins and endotoxin-coated polyethylene particles were introduced into rat femoral canals followed by intraperitoneal injection of pamidronate every other day. The treatment varied in the dose from 0 to 40 microg/kg and the duration of either 10 days or 6 weeks. Bilateral femurs were harvested after 6 weeks and examined by bone densitometer and MicroCT scan. Pamidronate increased the bone density of the left, unoperated femurs in a dose and duration dependent manner. Bone-titanium attachment significantly increased in all treatment groups compared to the control group. When pamidronate was administered for 10 days, the increase of bone-titanium attachment was significantly dose-dependent. However, when pamidronate was given for 6 weeks at 4 microg/kg, the bone-titanium attachment was significantly (p < 0.001) lower compared to the 10 day treatment of the same dose, although it was significantly higher than controls. Our results suggest that pamidronate effectively increase bone-titanium attachment even in the presence of endotoxin-coated polyethylene particles. However, long-term administration may reduce its efficacy.

Accumulation of LPS by polyethylene particles decreases bone attachment to implants.

Molecules absorbed on the surface of particulate wear debris may contribute to inflammatory reactions that lead to aseptic loosening of implants. Lipopolysaccharide (LPS), a bacterial endotoxin, can attach to many biomaterials and stimulate macrophages to secrete osteoclast-activating cytokines. We tested the adsorption of LPS by polyethylene particles in vitro and examined the biological effects of LPS absorption on bone remodeling around implants in vivo. Polyethylene particles were incubated in radiolabeled LPS solutions, and adsorption of LPS by the particles was quantified by radioassay. Because polyethylene particles are hydrophobic and less dense than water, they floated and clumped when incubated in a water solution of LPS, resulting in low adsorption of LPS. However, when particles were incubated in an ethanol solution of LPS, most of the LPS was adsorbed by the particles, and was resistant to washing with water. Triton X-100 (10%), however, effectively washed the LPS off the particles. In a rat model, the presence of polyethylene particles around the implant in the femoral canal decreased bone attachment to the implant at 6 weeks. Incubating the particles with LPS before implantation, or intermittent administration of LPS systemically, further decreased bone-implant attachment to similar extents, but had no effect on the bone density of the control side femurs. Our data indicate that polyethylene particles have high affinity for LPS, depending on many factors, especially the solvents of the LPS. Intermittent systemic administration of LPS affects bone remodeling but only occurs in the area containing polyethylene particles and titanium implants, supporting the hypothesis that the presence of polyethylene particles around implants can result in accumulation of LPS from exogenous sources. This may cause local levels of LPS that are high enough to affect bone remodeling around implants.

Titanium particles and surface-bound LPS activate different pathways in IC-21 macrophages.

It is still unknown if wear-debris particles themselves induce osteolysis or if they serve a functional role as receptors for ligands that incite an inflammatory response that ultimately leads to bone resorption. In this study, commercially pure titanium particles (cpTi) were subjected to a serial combination of different cleaning methods to remove Lipopolysaccharide (LPS) or were incubated in LPS solutions of known concentrations. Then, the response of the macrophage cell line IC-21 to the cleaned particles, LPS-bound Ti particles, and soluble LPS was examined. It was found that cleaned particles up to 1000 particles per cell did not stimulate macrophages to release Tumor necrosis factor-alpha (TNF-alpha) or Interleukin 6 (IL-6), but they significantly increased the release of Prostaglandin E(2) (PGE(2)) when the particle concentration was higher than 500 particles per cell. At one particle per cell, Ti particles bound with LPS stimulated the release of IL-6 and TNF-alpha by macrophages. The level of released cytokines was dependent on, and correlated with, the amount of LPS present on the particles. The macrophages were more sensitive to soluble LPS than to particle-bound LPS, and the simultaneous addition of cleaned Ti particles did not have additional effects on the effects of soluble LPS. This study shows evidence that, cpTi particles and LPS have distinct mechanisms of action on the IC-21 macrophages, but that both contribute to the development of an inflammatory response.

Hyperparathyroidism and the calcium paradox of aldosteronism.

BACKGROUND: Aldosteronism may account for oxi/nitrosative stress, a proinflammatory phenotype, and wasting in congestive heart failure. We hypothesized that aldosterone/1% NaCl treatment (ALDOST) in rats enhances Ca2+ and Mg2+ excretion and leads to systemic effects, including bone loss. METHODS AND RESULTS: At 1, 2, 4, and 6 weeks of ALDOST, we monitored Ca2+ and Mg2+ excretion, ionized [Ca2+]o and [Mg2+]o, parathyroid hormone and 1-antiproteinase activity in plasma, bone mineral density, bone strength, Ca2+ and Mg2+ content in peripheral blood mononuclear cells (PBMCs) and ventricular tissue, and lymphocyte H2O2 production. A separate group received spironolactone (Spiro), an aldosterone receptor antagonist. Age- and gender-matched unoperated and untreated rats served as controls. ALDOST induced a marked (P<0.05) and persistent rise in urinary and fecal Ca2+ and Mg2+ excretion, a progressive reduction (P<0.05) in [Ca2+]o and [Mg2+]o, and an elevation in parathyroid hormone (P<0.05) with a fall (P<0.05) in bone mineral density and strength. An early, sustained increase (P<0.05) in PBMC Ca2+ and Mg2+ was found, together with an increase (P<0.05) in tissue Ca2+. Plasma 1-antiproteinase activity was reduced (P<0.05), whereas lymphocyte H2O2 production was increased (P<0.05) at all time points. Spiro cotreatment attenuated (P<0.05) urinary and fecal Ca2+ and Mg2+ excretion, prevented the fall in [Ca2+]o and [Mg2+]o, rescued bone mineral density and strength, and prevented Ca2+ overloading of PBMCs and cardiomyocytes. CONCLUSIONS: In aldosteronism, Ca2+ and Mg2+ losses lead to a fall in [Ca2+]o and [Mg2+]o with secondary hyperparathyroidism and bone resorption. Ca2+ overloading of PBMCs and cardiac tissue leads to oxi/nitrosative stress and a proinflammatory phenotype.

Loss of bone minerals and strength in rats with aldosteronism.

Congestive heart failure (CHF) is a clinical syndrome with origins rooted in a salt-avid state largely mediated by effector hormones of the circulating renin-angiotensin-aldosterone system. Other participating neurohormones include catecholamines, endothelin-1, and arginine vasopressin. CHF is accompanied by a systemic illness of uncertain causality. Features include the appearance of oxidative/nitrosative stress and a wasting of tissues including bone. Herein we hypothesized that inappropriate (relative to dietary Na+) elevations in plasma aldosterone (Aldo) contribute to an altered redox state, augmented excretion of divalent cations, and in turn, a loss of bone minerals and strength. In uninephrectomized rats that received chronic Aldo and 1% NaCl treatment for 4-6 wk, we monitored plasma alpha1-antiproteinase activity, which is an inverse correlate of oxidative/nitrosative stress; plasma concentrations of ionized Mg2+ and Ca2+; urinary Mg2+ and Ca2+ excretion; and bone mineral composition and strength to flexure stress. Compared with controls, we found reductions in plasma alpha1-antiproteinase activity and ionized Mg2+ and Ca2+ together with persistently elevated urinary Mg2+ and Ca2+ excretion, a progressive loss of bone mineral density and content with reduced Mg2+ and Ca2+ concentrations, and a reduction in cortical bone strength. Thus the hypermagnesuria and hypercalciuria that accompany chronic Aldo-1% NaCl treatment contribute to the systemic appearance of oxidative/nitrosative stress and a wasting of bone minerals and strength.