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Calcific tendinitis, classified as enthesopathy, is a self-limiting disease that rarely involves the tendons of the gluteus maximus. We discuss a 52-year-old woman with a 1-year history of localized, reproducible posterolateral pain of her left hip, which was previously treated with steroid injection to her left greater trochanteric bursa without significant pain relief. Plain radiography and magnetic resonance imaging of the left hip revealed abnormal edema and calcifications at the insertion of the gluteus maximus tendon to the gluteal tuberosity, corresponding to the area of maximal tenderness on examination. This case report contributes to the growing medical literature for the rare presentation of calcific tendinitis of gluteus maximus insertion and reinforces the importance of the patient history, focused physical examination with special testing, and pertinent imaging for proper diagnosis and management.

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Calcinosis cutis is a quite unusual disease represented by abnormal accumulation of calcium salts in the skin and subcutaneous tissues. Repeated cutis calcinosis means recurrent calcium deposition in pre-existing areas. The case report illustrated the case of a 16-year-old female who had recurrent calcium deposits on the base of her right thumb. The patient initially had swelling at the base of her right thumb, which had been present for six months now. The patient described the dorsal solid mass on top of the thumb base, which was painful and had reduced thumb mobility. There was swelling that became painful, specifically located at the same site as the previous surgery, with thumb restriction and superadded infection at the metacarpophalangeal joint. Routine lab tests, including blood tests and rheumatologic and autoimmune work-ups, were normal. Plain radiographs and ultrasound examinations unveiled the characteristics of calcifications in the thumb tissues. A skin biopsy was done and the calcium deposits in subcutaneous tissue were confirmed, matching calcinosis cutis. The approach to the treatment of this condition entailed conservative measures. Some included physiotherapy to correct a flexion deformity, antibiotics, painkillers, and daily dressing. The patient was advised to follow up and to consider excision of the nodules. This case points out the clinical manifestations, investigations, and initial management of available strategies for recurrent calcinosis cutis. Further studies and long-term follow-up are necessary to determine the optimal treatment approaches and outcomes for this rare condition.

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BACKGROUND: Bone morphogenetic proteins (BMPs) are used in clinical practice for stimulation of bone formation, but often evoke serious complications. Recent studies demonstrated that BMPs involved in early stages of bone formation are species specific. In cattle dominate BMP7, growth differentiation factor 5 (GDF5) and NEL-like protein 1 (NELL1) while in rats BMP2, BMP5 and BMP6. The purpose of the study was to compare the action of the species specific BMPs on the osteoprogenitor cells. Thus, rat osteoprogenitor cells were exposed to one BMP in a high dose and three of them at 1/3 of the former. MATERIALS AND METHODS: Isolated rat osteoprogenitor cells were treated in culture with different concentrations of BMP2, BMP5 and BMP6 or with lower concentration of combinations of these cytokines. Activity of alkaline phosphatase, calcium deposition and mRNA level for transcription factor SP7 (osterix) and tissue non-specific alkaline phosphatase (TNAP) served as indicators of BMPs effect. RESULTS: BMPs stimulated all studied parameters in comparison with control cultures, but no statistically significant differences were observed between the action of a large dose of one cytokine and a combination of cytokines given at lower concentrations. CONCLUSIONS: Three BMPs used in a low dose exert similar effect as the one used at high dose. Since the BMPs stimulate different receptors and activate different signaling pathways the use of the mixture of properly chosen BMPs at low concentration may give better results than the single one at high concentration and may avoid untoward effects.

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Objective: To describe acute calcareous corneal degeneration as a complication of chronic graft-versus-host disease. Materials and methods: Clinical case and review of the literature. Results: We presented a case of bilateral acute calcareous corneal degeneration in a patient with chronic graft-versus-host disease. Conclusions: Chronic graft-versus-host disease (cGVHD) occurs in 50-70% of bone marrow transplantation patients, the most frequent ocular complication being keratoconjunctivitis sicca (KCS). Calcareous corneal degeneration is a type of calcium deposition that can be secondary to chronic ocular inflammation or dry eye, but there are few cases reported of acute calcareous corneal degeneration and recurrent perforation in cGVHD. Abbreviations: GVHD = Chronic graft-versus-host disease, aGVHD = Acute graft-versus-host disease, cGVHD = Chronic graft-versus-host disease, KCS = Keratoconjunctivitis sicca, PKP = Penetrating keratoplasty, AMT = Amniotic membrane transplantation, PRGF = Plasma rich in growth factors, OD = Right eye, OS = Left eye.

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Guided bone regeneration (GBR) comprehends the application of membranes to drive bone healing and to exclude non-osteogenic tissues from interfering with bone regeneration. However, the membranes may be exposed to bacterial attack, with the risk of failure of the GBR. Recently, an antibacterial photodynamic protocol (ALAD-PDT) based on a gel with 5% 5-aminolevulinic acid incubated for 45 min and irradiated for 7 min by a LED light at 630 nm, also showed a pro-proliferative effect on human fibroblasts and osteoblasts. The present study hypothesized that the functionalization of a porcine cortical membrane (soft-curved lamina, OsteoBiol) with ALAD-PDT might promote its osteoconductive properties. TEST 1 aimed to verify the response of osteoblasts seeded on lamina with respect to the plate surface (CTRL). TEST 2 aimed to investigate the effects of ALAD-PDT on the osteoblasts cultured on the lamina. SEM analyses were performed to study the topographical characteristics of the membrane surface, the adhesion, and the morphology of cells at 3 days. The viability was assessed at 3 days, the ALP activity at 7 days, and calcium deposition at 14 days. Results showed the porous surface of the lamina and the increase in cell attachment of osteoblasts with respect to controls. The proliferation, the ALP, and bone mineralization activity of osteoblasts seeded on lamina resulted in being significantly higher (p < 0.0001) than controls. Results also showed an additional significative enhancement (p < 0.0001) in the proliferative rate in ALP and calcium deposition after applying ALAD-PDT. In conclusion, the functionalization of the cortical membranes cultured with osteoblasts with the ALAD-PDT improved their osteoconductive properties.

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The study in this paper presents a new material that was produced as a thin film by the Pulsed Laser Deposition technique (PLD) using a 532 nm wavelength and 150 mJ/pulse laser beam on the hemp stalk as target. The analyses performed by spectroscopic techniques (Fourier Transform Infrared Spectroscopy-FTIR, Laser-Induced Fluorescence Spectroscopy-LIF, Scanning Electron Microscopy coupled with Energy Dispersive X-ray-SEM-EDX, Atomic Force Microscopy-AFM and optical microscope) evidenced that a biocomposite consisting of lignin, cellulose, hemicellulose, waxes, sugars and phenolyc acids p-coumaric and ferulic, similar to the hemp stalk target was obtained. Nanostructures and aggregated nanostructures of 100 nm to 1.5 µm size were evidenced. Good mechanical strength and its adherence to the substrate were also noticed. It was noticed that the content in calcium and magnesium increased compared to that of the target from 1.5% to 2.2% and from 0.2% to 1.2%, respectively. The COMSOL numerical simulation provided information on the thermal conditions that explain phenomena and processes during laser ablation such as C-C pyrolisis and enhanced deposition of calcium in the lignin polymer matrix. The good gas and water sorption properties due to the free OH groups and to the microporous structure of the new biocomposite components recommends it for studies for functional applications in medicine for drug delivery devices, filters in dialysis and for gas and liquid sensors. Functional applications in solar cells windows are also possible due to the conjugated structures of the contained polymers.

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BACKGROUND: Vitamin D receptor (VDR) is critical for mineral and bone homeostasis since it plays an essential role in the osteoblast differentiation of bone marrow mesenchymal stem cells (BM-MSCs). Hydroxysafflor yellow A (HSYA) has the potential to promote bone mineralization and inhibit bone resorption, while its detailed mechanism needs to be elaborated. OBJECTIVE: This study intends to explore the action of HSYA on the proliferation and differentiation of BM-MSC and the underlying mechanism. METHODS: Different concentrations of HSYA to BM-MSC and CCK-8, and EdU were used to detect cell viability and proliferation. The alkaline phosphatase (ALP) was used to observe the differentiation ability of BM-MSC osteoblasts. The calcium uptake and mineralization of osteoblast-like cells were observed by alizarin red staining. The level of calcium ion uptake in cells was detected by flow cytometry. AutoDock was performed for molecular docking of HSYA to VDR protein. Immunofluorescence and western blotting were performed to detect the expression of VDR expression levels. Finally, the effect of VDR was verified by a VDR inhibitor. RESULTS: After treatment with HSYA, the proliferation and calcium uptake of BM-MSC were increased. The level of ALP increased significantly and reached its peak on the 12th day. HSYA promoted calcium uptake and calcium deposition, and mineralization of osteoblasts. The western blotting and immunofluorescence showed that HSYA increased the expression of VDR in the osteoblast-like cell's nucleus and upregulated Osteocalcin, S100 calcium-binding protein G, and CYP24A1. In addition, HYSA treatment increased the expression of osteopontin and the synthesis of osteogenic proteins, such as Type 1 collagen. After the addition of the VDR inhibitor, the effect of HSYA was weakened. CONCLUSION: HSYA could significantly promote the activity and proliferation of osteoblasts and increase the expression level of VDR in osteoblasts. HSYA may also improve calcium absorption by osteoblasts by regulating the synthesis of calciumbinding protein and vitamin D metabolic pathway-related proteins.

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Fahr's syndrome, recently named "primary brain calcification," is a rare disorder with a variable clinical presentation ranging from behavioral changes to seizures. It can be idiopathic or have multiple causes, hypoparathyroidism the most frequent. In the current coronavirus 2019 (COVID-19) pandemic, these electrolyte imbalances have acquired importance, and there has been a correlation between the lowest serum calcium levels and severe COVID-19 disease. It is known that calcium accomplishes many normal physiologic functions. We present a case of a 63-year-old woman who arrived at the emergency room with a fever of 10-day duration, odynophagia, dry cough, dyspnea, and drowsiness. Upon her arrival, computed tomography of the brain and chest was performed, showing areas of calcification in the basal nuclei and infiltrates with a ground-glass pattern, respectively. In addition, laboratory studies were conducted in which hypocalcemia and hyperphosphatemia stand out. Furthermore, a positive result was obtained from acute Respiratory Syndrome Coronavirus 2 (SARS-COV2) from bronchial secretion. According to the clinical presentation data in the imaging and laboratory studies, Fahr's syndrome and COVID-19 pneumonia were diagnosed. We consider evaluating electrolyte imbalances at case presentations essential and continuously monitoring them. Appropriate and prompt corrections were achieved in patients with hypoparathyroidism history and severe COVID-19 disease. This case shows the vital collaboration between endocrinologists and other physicians that care for patients with COVID-19 infection.

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Calcium transportation and homeostasis are essential for marine bivalves to maintain basic metabolism and build their shells. Calmodulin-like proteins (CaLPs) are important calcium sensors and buffers and can respond to ocean acidification (OA) in marine calcifiers. However, no further study of their physiological function in calcium metabolism under elevated CO2 has been performed. Here, we identified a novel CaLP (designated CgCaLP) in the Pacific oyster Crassostrea gigas and demonstrated its participation in the calcification process: the mRNA expression level of CgCaLP peaked at the trochophore larval stage and remained high at stages when shells were shaped; the mRNA and protein of CgCaLP were more highly expressed in mantle tissue than in other tissues. Under elevated CO2 levels, the protein expression level of CgCaLP in hemocytes increased, while in contrast, significantly decreased protein levels were detected in gill and mantle tissues. Shell dissolution caused the imbalance of calcium in hemocytes and decreased calcium absorption and transportation demand in gill and mantle tissues, inducing the molecular function allocation of CgCaLP under CO2 exposure. Despite the decreased protein level in mantle tissue, CgCaLP was found to translocate to outer mantle epithelium (OME) cells where condensed calcium-rich deposits (CRDs) were detected. We further demonstrated that CgCaLP mRNA and protein expression levels could respond to seawater Ca2+ availability, suggesting that the calcium deposition capacity of oysters might be enhanced to fight against shell dissolution problems and that CgCaLP might serve as an essential participator of the process. In summary, CgCaLP might enhance calcium deposition under CO2 exposure and thus play a significant and flexible molecular function involved in a compensation strategy of oysters to fight against the acidified ocean.

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Background: Serum uric acid (SUA) is suspected to be associated with atherosclerosis and calcium deposition in atherosclerosis is known to related poor prognosis, yet there is no cohort study on the aged in China. We aimed to investigate the relationships between SUA levels and coronary calcium deposition in the middle-aged and elderly populations in China. Methods: A total of 326 participants between the ages of 50 and 85 who had undergone a coronary CT scan in 2015 at the Huadong Hospital Affiliated to Fudan University (Shanghai, China) were included in this study. Univariate and multivariate binary logistic regression was performed to analyze the correlation between SUA levels and coronary artery calcium score (CACS). The changes in CACS during a five-year follow-up were analyzed through Kaplan-Meier survival and binary cox regression analysis. An observational study was done on another 104 asymptomatic middle-aged and elderly patients to compare relative mRNA expressions of proinflammatory factors in peripheral blood mononuclear cells (PBMCs) from 104 subjects. Results: Based on the first year of follow-up data analysis, the elevation of SUA levels (P<0.001) is an independent risk factor for the increase of CACS after coordinating the confounding factors. According to five-year follow-up data, cox regression analysis proved that SUA was a risk factor for CACS (HR =5.86, P<0.001). The mRNA expression of IL-6 and CXCL8 in the HUA and HUA patients with CAC (HUA-CAC) groups was significantly higher than that in the normal control (NC) and coronary calcium deposition (CAC) groups. Conclusion: Taken together, the findings in this study indicate that high SUA levels (P<0.001) are an independent risk factor for CACS and elevated SUA levels increase the risk of developing coronary calcium deposition among middle-aged and old people in the Chinese population, which may be related to an increase of pro-inflammatory cytokines in the PBMCs.

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Aims: Vascular calcification is a common clinical complication of chronic kidney disease (CKD), atherosclerosis (AS), and diabetes, which is associated with increased cardiovascular morbidity and mortality in patients. The transdifferentiation of vascular smooth muscle cells (VSMCs) to an osteochondrogenic phenotype is a crucial step during vascular calcification. The transcription factor CCAAT/enhancer-binding protein alpha (C/EBPα) plays an important role in regulating cell proliferation and differentiation, but whether it regulates the calcification of arteries and VSMCs remains unclear. Therefore, this study aims to understand the role of C/EBPα in the regulation of vascular calcification. Methods and Results: Both mRNA and protein expression levels of C/EBPα were significantly increased in calcified arteries from mice treated with a high dose of vitamin D3 (vD3). Upregulation of C/EBPα was also observed in the high phosphate- and calcium-induced VSMC calcification process. The siRNA-mediated knockdown of C/EBPα significantly attenuated VSMC calcification in vitro. Moreover, C/EBPα depletion in VSMCs significantly reduced the mRNA expression of the osteochondrogenic genes, e.g., sex-determining region Y-box 9 (Sox9). C/EBPα overexpression can induce SOX9 overexpression. Similar changes in the protein expression of SOX9 were also observed in VSMCs after C/EBPα depletion or overexpression. In addition, silencing of Sox9 expression significantly inhibited the phosphate- and calcium-induced VSMC calcification in vitro. Conclusion: Findings in this study indicate that C/EBPα is a key regulator of the osteochondrogenic transdifferentiation of VSMCs and vascular calcification, which may represent a novel therapeutic target for vascular calcification.

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OBJECTIVE: To investigate the effect of Bax inhibitor-1(BI-1)on calcification of vascular smooth muscle cells(VSMCs). METHODS: VSMCs were isolated from the thoracic aorta of SD rats.VSMCs or BI-1-overexpressing VSMCs(transfected with a BI-1-overexpressing plasmid) were cultured in normal medium or calcified medium containing ß-glycerophosphate and calcium chloride, and the cell calcification was examined with Alizarin red staining.Enzyme-linked immunosorbent assay was used to determine the intracellular calcium content and alkaline phosphatase activity.The expression levels of Runt-related transcription factor 2 (RUNX2), bone morphogenetic protein 2 (BMP-2) and caspase-3 were detected with Western blotting. RESULTS: After 14 days of culture in the calcified medium, the VSMCs showed significantly reduced expression of BI-1 protein(P=0.001).BI-1 overexpression in the VSMCs caused a significant reduction of calcium level and alkaline phosphatase activities(P=0.0006) and lowered the expression levels of RUNX2 and BMP-2 (P=0.0001) in the cells.The VSMCs with induced calcification exhibited a significantly increased apoptosis rate, but BI-1 overexpression obviously inhibited VSMC apoptosis in the calcified medium (P=0.0003). CONCLUSION: BI-1 may attenuate vascular calcification by inhibiting calcium deposition, osteogenic differentiation and apoptosis.

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Platelet-rich fibrin (PRF) is a natural fibrin meshwork material with multiple functions that are suitable for tissue engineering applications. PRF provides a suitable scaffold for critical-size bone defect treatment due to its platelet cytokines and rich growth factors. However, the structure of PRF not only promotes cell attachment but also, due to its density, provides a pool for cell migration into the PRF to facilitate regeneration. In our study, we used repeated freeze drying to enlarge the pores of PRF to engineer large-pore PRF (LPPRF), a type of PRF that has expanded pores for cell migration. Moreover, a biodegradable Mg ring was used to provide stability to bone defects and the release of Mg ions during degradation may enhance osteoconduction and osteoinduction. Our results revealed that cell migration was more extensive when LPPRF was used rather than when PRF was used and that LPPRF retained the growth factors present in PRF. Moreover, the Mg ions released from the Mg ring during degradation significantly enhanced the calcium deposition of MC3T3-E1 preosteoblasts. In the present study, a bone substitute comprising LPPRF combined with a Mg ring was demonstrated to have much potential for critical-size bone defect repair.

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For birds, the uterus is an important part for eggshell mineralization, and the establishment of the endometrial epithelial cell (EEC) model was beneficial to the study of uterine function. This study was conducted to establish a culture model of primary EECs of laying hens and explore the effects of zinc on primary EEC proliferation, zinc transport, and calcium deposition in vitro. The EECs were isolated and cultured via type I collagenase digestion, and in the logarithmic phase during 2-5 days, and then reached the plateau phase on the 7th day of inoculation. Results showed that the proliferation of EECs treated by 50 µM ZnSO4 or zinc-methionine (Zn-Met) were markedly promoted at 24-h or 48-h treating time (P < 0.05). In later experiments, the EECs were divided into three groups, involving a control group (no zinc treated), ZnSO4 group (50 µM zinc treated) and a Zn-Met group (50 µM zinc treated). Results showed the relative fluorescence intensity of Ca2+ in the Zn-Met group was significantly higher than that in the control group (P < 0.05). As for zinc transporters, it was only observed that mRNA levels of metallothionein (MT) in EECs showed a significant difference (P < 0.05) between the Zn-Met group and the control. In conclusion, the EECs of laying hens isolated by scraping and digested collagenase I were with better adherent growth. Moreover, Zn-Met can increase intracellular Ca2+ concentration and upregulate expressions of MT mRNA in the EECs of laying hens.

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BACKGROUND: Intraocular ossification is an uncommon calcium deposition process associated with trauma, chronic inflammation, tumor, and long-standing retinal detachment. This is the first reported extensive intraocular bone formation associated with silicone oil. CASE PRESENTATION: A 30-year-old Han Chinese man came to us with complaint of red, painful blind right eye. He had a history of ocular trauma, retinal detachment, and two failed retinal reattachment surgeries with silicone oil left in the eye. On examination, conjunctiva congestion, band keratopathy, silicone oil emulsification, and limbus neovascularization were found. B-scan ultrasound and computed tomography scanning demonstrated retinal detachment and calcification of the eyeball wall. Histopathological analysis indicated ossification overlying the choroid. Evisceration was finally operated to relieve the pain. CONCLUSION: The retention of silicone oil in the eye probably accelerates the ossification. Timely silicone oil removal and evisceration should be recommended if necessary for phthisis bulbi.

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Herein, K-OMS-2 catalysts with different levels of Ca2+ loading were synthesized to investigate the influence of Ca2+ deposition on the catalytic oxidation of CB. The micromorphology, redox ability, oxygen species, and surface acidity of the prepared catalysts were analyzed via SEM, HR-TEM, H2-TPR, O2-TPD, Py-IR, and GC-MS. After the Ca addition, CB conversion on the catalyst was achieved at <220 °C and the polychlorinated by-product yield decreased significantly. Although CaCO3 formation on the catalyst surface from Ca2+ caused a decline in number and fluidity of the surface lattice oxygen species, no significant impact on surface adsorbed oxygen species content was observed. Furthermore, CaCO3 reacted with dissociated chlorine species (HCl and polychlorinated products generated in situ), which inhibited chlorine poisoning of the active phase.

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A series of biocompatible high-porosity (up to 72.4%) TiZr-based porous bulk metallic glass (BMG) scaffolds were successfully fabricated by hot pressing a mixture of toxic element-free TiZr-based BMG powder and an Al particle space holder. The morphology of the fabricated scaffolds was similar to that of human bones, with pore sizes ranging from 75 to 250 µm. X-ray diffraction patterns and transmission electron microscopy images indicated that the amorphous structure of the TiZr-based BMG scaffolds remained in the amorphous state after hot pressing. Noncytotoxicity and extracellular calcium deposition of the TiZr-based BMG scaffolds at porosities of 32.8%, 48.8%, and 64.0% were examined by using the direct contact method. The results showed that the BMG scaffolds possess high cell viability and extracellular calcium deposition with average cell survival and deposition rates of approximately 170.1% and 130.9%, respectively. In addition, the resulting TiZr-based BMG scaffolds exhibited a considerable reduction in Young's moduli from 56.4 to 2.3 GPa, compressive strength from 979 to 19 MPa, and bending strength from 157 MPa to 49 MPa when the porosity was gradually increased from 2.0% to 72.4%. Based on the aforementioned specific characteristics, TiZr-based BMG scaffolds can be considered as potential candidates for biomedical applications in the human body.

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In this study, we used various proteinases to investigate the effect of whey protein hydrolysates on proliferation and differentiation of MC3T3-E1 osteoblasts. To confirm hydrolysis of the whey protein hydrolysates, the yield and α-amino acid content were determined. Since osteogenic cell activity is an important factor in osteogenesis, we evaluated the proliferation of osteogenic cells by measuring 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide and alkaline phosphatase (ALP) activity. To analyze bone matrix formation, we identified calcium deposition by staining with Alizaline red-S. The free amino acid content was significantly higher in the whey protein hydrolysates prepared using Protamex, Flavourzyme, and Alcalase than in the control. When cells were treated with 500 µg/mL of whey protein hydrolysates prepared using Protamex and Alcalase, cell proliferation increased by 120% and 130%, respectively, compared with the control group. In addition, ALP activity was significantly higher following treatment with 500 µg/mL of whey protein hydrolysates prepared using Protamex and Alcalase (142.61% and 135.06%, respectively; P<0.05). Furthermore, when treated with 125 µg/mL of the same hydrolysates, the rate of calcium deposition increased significantly to 157.56% compared with the control group (P<0.05). Therefore, our results suggest that whey protein hydrolysates prepared using Protamex and Alcalase may have more beneficial effects on osteoblast proliferation and bone health than those prepared using other proteolytic enzymes.