Long-Term Safety of Bone Regeneration Using Autologous Stromal Vascular Fraction and Calcium Phosphate Ceramics: A 10-Year Prospective Cohort Study.

This prospective cohort study aimed to assess long-term safety, dental implant survival, and clinical and radiological outcomes after maxillary sinus floor elevation (MSFE; lateral window technique) using freshly isolated autologous stromal vascular fraction (SVF) combined with calcium phosphate ceramics. All 10 patients previously participating in a phase I trial were included in a 10-year follow-up. They received either ß-tricalcium phosphate (ß-TCP; n = 5) or biphasic calcium phosphate (BCP; n = 5) with SVF-supplementation on one side (study). Bilaterally treated patients (6 of 10; 3 ß-TCP, 3 BCP) received only calcium phosphate on the opposite side (control). Clinical and radiological assessments were performed on 44 dental implants at 1-month pre-MSFE, and 0.5- to 10-year post-MSFE. Implants were placed 6 months post-MSFE. No adverse events or pathology was reported during a 10-year follow-up. Forty-three dental implants (98%) remained functional. Control and study sides showed similar peri-implant soft-tissue quality, sulcus bleeding index, probing depth, plaque index, keratinized mucosa width, as well as marginal bone loss (0-6 mm), graft height loss (0-6 mm), and graft volume reduction. Peri-implantitis was observed around 6 implants (control: 4; study: 2) in 3 patients. This study is the first to demonstrate the 10-year safety of SVF-supplementation in MSFE for jawbone reconstruction. SVF-supplementation showed enhanced bone regeneration in the short term (previous study) and led to no abnormalities clinically and radiologically in the long term.

The Antioxidant Supplementation with Filipendula ulmaria Extract Attenuates the Systemic Adverse Effects of Nanosized Calcium Phosphates in Rats.

The aim of this study was to investigate and compare the systemic toxicity of three nanosized calcium phosphates (CaPs): hydroxyapatite (HA), tricalcium phosphate (TCP), and amorphous calcium phosphate (ACP) in rats. Since those metallic compounds are widely used as bone replacement materials, including their use in oral surgery, CaPs were applied (per os) equimollary (17.8 mg/kg, 11 mg/kg, and 9.65 mg/kg b.w., respectively) for 30 days in order to mimic the previously described release rate from dental composites. Also, we employed antioxidant supplementation with Filipendula ulmaria (FU) extract. All the applied CaPs significantly increased serum calcium, triglycerides, LDL, and LDH, while serum levels of testosterone and LH declined, with no alterations in the liver enzymes. The evaluation of oxidative stress markers (in the liver, kidney, and testicle) showed an increase in TBARS values, while SOD and CAT activities and GSH levels were significantly reduced. The relative gene expression of Bax and Bcl-2 was shifted to proapoptotic action, accompanied by intense characteristic histological changes in architecture in all investigated organs. The toxic effects were most prominent in groups treated by ACP. FU administration attenuated the majority of nanosized CaP-induced adverse effects, thus recommending this therapeutic approach to minimize nano-CaP systemic toxicities.

Modifications to advanced Core decompression for treatment of Avascular necrosis of the femoral head.

BACKGROUND: "Advanced Core Decompression" (ACD) is a new technique for treatment of osteonecrosis of the femoral head (ONFH) that includes removal of the necrotic tissue using a percutaneous expandable reamer followed by refilling of the drill hole and the defect with an injectable, hard-setting, composite calcium sulphate (CaSO4)-calcium phosphate (CaPO4) bone graft substitute. As autologous bone has been shown to be superior to all other types of bone grafts, the aim of the study is to present and evaluate a modified technique of ACD with impaction of autologous bone derived from the femoral neck into the necrotic defect. METHODS: A cohort of patients with an average follow-up of 30.06 months (minimum 12 months) was evaluated for potential collapse of the femoral head and any reasons that led to replacement of the operated hip. Only patients in stages 2a to 2c according to the Steinberg classification were included in the study. RESULTS: In 75.9% the treatment was successful with no collapse of the femoral head or conversion to a total hip replacement. Analysis of the results of the different subgroups showed that the success rate was 100% for stage 2a lesions and 84.6% respectively 61.5% for stages 2b and 2c lesions. CONCLUSIONS: Previous studies with a comparable follow-up reported less favourable results for ACD without autologous bone. Especially in stages 2b and 2c the additional use of autologous bone has a positive effect. In comparison to other hip-preserving techniques, the modified ACD technique is a very promising and minimally invasive method for treatment of ONFH. TRIAL REGISTRATION: German clinical trials register ( DRKS00011269 , retrospectively registered).

Calcium phosphate: a substitute for aluminum adjuvants?

INTRODUCTION: Calcium phosphate was used as an adjuvant in France in diphtheria, tetanus, pertussis and poliomyelitis vaccines. It was later completely substituted by alum salts in the late 80's, but it still remains as an approved adjuvant for the World Health Organization for human vaccination. Area covered: Thus, calcium phosphate is now considered as one of the substances that could replace alum salts in vaccines. The aim of this paper is to draw a review of existing data on calcium phosphate as an adjuvant in order to bring out the strengths and weaknesses for its use on a large scale. Expert commentary: Calcium phosphate is a compound naturally present in the organism, safe and already used in human vaccination. Beyond comparisons with the other adjuvants, calcium phosphate represents a good candidate to replace or to complete alum salts as a vaccine adjuvant.

Calcium Supplementation Does Not Contribute to Constipation in Healthy Women.

PURPOSE: It is commonly suggested that calcium supplementation contributes to constipation; however, little research has explored the effects of calcium supplementation on gut motility. METHODS: In an 8-week, randomized, double-blind, crossover pilot study, healthy females (n = 27, aged 43.0 ± 10.6 years) received a split dose of 500 mg/d of elemental calcium from calcium carbonate or calcium phosphate each for 2 weeks, after a 2-week baseline and separated by a 2-week washout. Participants completed daily questionnaires of stool frequency, Bristol Stool Form Scale (BSFS), and supplement intake compliance. RESULTS: There were no differences among periods. Mean ± SE stool frequency averaged 1.3 ± 0.1 stools/d in each period. Participants reported 34%, 34%, 37%, and 29% of stools were indicative of slow transit or constipation (BSFS of 1 or 2) during baseline, calcium carbonate, calcium phosphate, and washout periods, respectively. Participants also reported from 6% to 10% of stools as fast transit or diarrhea (BSFS of 6 or 7) during the periods. CONCLUSION: This study suggests that neither calcium carbonate nor calcium phosphate, providing 500 mg/d of calcium, affects stool frequency or form. Although stool frequency was normal, the healthy females participating in the study experienced stools indicating slow (constipation) and fast (diarrhea) transit.

Evaluation of four biodegradable, injectable bone cements in an experimental drill hole model in sheep.

Four cement applications were tested in this investigation. Two dicalcium phosphate dihydrate (DCPD-brushite) hydraulic cements, an apatite hydraulic fiber loaded cement, and a calcium sulfate cement (Plaster of Paris) were implanted in epiphyseal and metaphyseal cylindrical bone defects in sheep. The in vivo study was performed to assess the biocompatibility and bone remodeling of four cement formulations. After time periods of 2, 4, and 6 months, the cement samples were clinically and histologically evaluated. Histomorphometrically, the amount of new bone formation, fibrous tissue, and bone marrow and the area of remaining cement were measured. In all specimens, no signs of inflammation were detectable either macroscopically or microscopically. Cements differed mainly in their resorption time. Calcium sulfate was already completely resorbed at 2 months and showed a variable amount of new bone formation and/or fibrous tissue in the original drill hole over all time periods. The two DCPD cements in contrast were degraded to a large amount at 6 months, whereas the apatite was almost unchanged over all time periods.

Effect of bamboo salt-NaF dentifrice on enamel remineralization.

PURPOSE: To evaluate the laboratory remineralization effects of a dentifrice with bamboo salt and NaF on artificial caries-like enamel lesions, at both the surface and deep areas. METHODS: Early dental caries lesions were formed by treating bovine enamel samples for 48 hours at 37 degrees C with a demineralization solution (pH 5.0) containing 0.1 M lactic acid, 0.2% Carbopol 907, and 50% saturated calcium phosphate tribasic. pH cycling was then performed by immersing the samples in dentifrice slurry for 2 minutes every 8 hours per day, and in demineralization solution for 4 hours and mixed saliva for the remaining time period. The mixed saliva consisted of 50% human saliva and 50% artificial saliva. The surface hardness and the level of mineral surface alterations were analyzed using a hardness tester and transversal microradiography, among negative control (fluoride free), positive control (sodium fluoride 1100 ppm, Crest Cavity Protection), and test dentifrice (3.0% bamboo salt with sodium fluoride 1,000 ppm) groups. RESULTS: Test and positive control groups significantly increased the level of the surface hardness and decreased mineral loss of the artificial caries-like enamel lesions compared to the negative control (P<0.05). The test dentifrice also significantly decreased the lesion depth compared to the other two groups (P<0.05).

Calcium carbonate suppresses haem toxicity markers without calcium phosphate side effects on colon carcinogenesis.

Red meat intake is associated with an increased risk of colorectal cancer. We have previously shown that haemin, Hb and red meat promote carcinogen-induced preneoplastic lesions, aberrant crypt foci (ACF), in the colon of rats. We have also shown that dietary calcium phosphate inhibits haemin-induced promotion and normalises faecal lipoperoxides and cytotoxicity. Unexpectedly, high-calcium phosphate control diet-fed rats had more preneoplastic lesions in the colon than low-Ca control diet-fed rats. The present study was designed to find a Ca supplementation with no adverse effect, by testing several doses and types of Ca salts. One in vitro study and two short-term studies in rats identified calcium carbonate as the most effective Ca salt to bind haem in vitro and to decrease faecal biomarkers previously associated with increased carcinogenesis: faecal water cytotoxicity and thiobarbituric acid-reactive substances. A long-term carcinogenesis study in dimethylhydrazine-injected rats demonstrated that a diet containing 100 µmol/g calcium carbonate did not promote ACF, in contrast with a previously tested calcium phosphate diet. The results suggest that calcium carbonate, and not calcium phosphate, should be used to reduce haem-associated colorectal cancer risk in meat eaters. They support the concept that the nature of the associated anion to a protective metal ion is important for chemoprevention.

Incorporation of methotrexate in calcium phosphate cement: behavior and release in vitro and in vivo.

The application of antibiotic-loaded calcium phosphate bone cement in the treatment and prevention of osteomyelitis suggests that calcium phosphate cement could also be used as an anticancer drug carrier to reduce the local recurrence of bone tumors and systemic toxicities of chemotherapy. We added 0 to 400 mg of methotrexate to 40 g cement, with final methotrexate concentrations of 0% to 1% (weight/weight). The setting times, mechanical properties, microstructures, and in vitro methotrexate release kinetics of these methotrexate-calcium phosphate bone cement specimens were evaluated, along with in vivo methotrexate release kinetics in 24 rabbits. Methotrexate did not significantly alter the cement setting time. The compressive and tensile strengths of the methotrexate-calcium phosphate bone cement specimens were significantly less when 400 mg methotrexate was used, compared to control samples without methotrexate addition. Nevertheless, the cement remained compliant with the minimum requirements for clinical application. The scanning electron microscopy micrographs showed that the basic crystal structure did not alter. The methotrexate release kinetics in vitro and in vivo confirmed that methotrexate-calcium phosphate bone cement was a monolithic matrix system, with a burst effect in the initial stage and a sudden drop thereafter. Drug delivery in vivo was faster than in vitro. We estimated that the incorporated methotrexate could be continuously released over 2 to 4 months at a higher than minimum concentration. The methotrexate had no apparent toxicity on the host rabbits, even at the highest dose of methotrexate tested. The methotrexate-calcium phosphate bone cement system may be a potentially effective therapy for bone tumors in humans.

Fabrication and evaluation of a new composite composed of tricalcium phosphate, gelatin and chi-li-saan as a bone substitute.

The purpose of this study was to prepare and evaluate the feasibility and biocompatibility of a new composite as a bone substitute. The new composite (GTGC) was mainly composed of tricalcium phosphate ceramics and gelatin to which chi-li-saan, a Chinese medicinal remedy was added. The GTGC composite was manually packed into cylindrical Teflon molds, dried overnight in an oven and sterilized by gamma-ray prior to use. Mature New Zealand rabbits, weighting 3-3.5 kg, underwent full-thickness excision of the parietal bone. In the experimental group, bone defects of 12 animals were filled with the GTGC composites and another 12 unreconstructed rabbits were considered as controls. Three rabbits were examined for each group in every time period at 2, 4, 8 and 12 weeks after operation. There was no evidence of adverse tissue reaction to the GTGC composite. In addition, examination with light and fluorescent microscopy revealed a significantly greater amount of new bone ingrowth in the GTGC group at the same implantion time as compared with the controls. Therefore, the GTGC composite could serve as a useful substitute when repairing bone defects.

Booster vaccination against diphtheria and tetanus in man. Comparison of calcium phosphate and aluminium hydroxide as adjuvants--II.

Diphtheria and tetanus antibody levels were measured before and four weeks after booster vaccination of 313 Danish military recruits participating in a clinical trial to compare aluminium hydroxide and calcium phosphate as adjuvants in diphtheria-tetanus vaccines (DT). Twenty-eight percent of the men had a diphtheria pre-vaccination content below a protective level of 0.01 IU ml-1. The calcium phosphate adsorbed vaccine showed the highest efficacy for both antigens. Adverse reactions were rare but more frequent in the calcium group than in the aluminium group. No correlation was found between pre- or post-vaccination levels and adverse reactions and both vaccines gave rise to specific IgE formation. The results show that calcium phosphate is more effective but not a safer alternative to aluminium hydroxide when compared in vaccines containing 1.0 mg ml-1 of Ca or of Al.

Laying hens respond differently to high dietary levels of phosphorus in monobasic and dibasic calcium phosphate.

A series of experiments were conducted to determine the relative acidogenicities of dietary chloride, phosphorus, and sulfate and their effects on shell quality. Calcium salts of these anions were added to a corn-soybean meal diet to provide supplemental levels of 150 to 450 meq/kg diet chloride, 150 to 600 meq/kg diet monobasic phosphate, 150 to 1,200 meq/kg diet dibasic phosphate, and 150 to 1,200 meq/kg diet sulfate. The results indicated that laying hens can safely tolerate up to 200 meq/kg diet supplemental chloride (.95% total chloride in the diet) and up to 450 to 600 meq/kg diet supplemental sulfate (2.16 to 2.88% added sulfate). Levels higher than these adversely affected shell quality and altered blood acid-base balance. Laying hens responded quite differently to phosphorus when it was supplied as monobasic or dibasic phosphate. Supplemental levels of 900 to 1,200 meq/kg diet dibasic phosphate (1.95 to 2.41% total phosphorus) did not have an adverse effect on performance and up to a supplemental level of 450 meq/kg diet (1.25% total phosphorus) did not affect shell quality. In contrast, performance was seriously impaired by a supplemental level of 150 meq/kg diet monobasic phosphate (1.02% total phosphorus in the diet). The results further indicated that phosphate in monobasic (but not in dibasic) form is a strong acidogenic anion and that most of its adverse effect on performance and shell quality can be ameliorated by alkaline salts of sodium or potassium bicarbonate. The relative acidogenicities of calcium salts of these acidic anions were as follows: phosphate (monobasic) > chloride > sulfate > phosphate (dibasic).

Calcium phosphate crystal occlusion of central venous catheters used for total parenteral nutrition in infants and children: prevention and treatment.

Calcium phosphate crystal occlusion is a complication occasionally encountered with long-term indwelling Silastic central venous catheters used for total parenteral nutrition (TPN) in infants and children. These occluded catheters are usually treated by removal. We have successfully treated six patients who experienced seven episodes of calcium phosphate crystal central venous catheter occlusion by irrigating their catheters with a hydrochloric (HCl) acid heparin solution. Although temporary febrile reactions occurred in three cases (42%), no serious complications were encountered. An average of 46 catheter-days per patient episode were preserved. By paying close attention to the calcium and phosphate concentrations in a patient's TPN solution, the clinician can minimize the risk of calcium phosphate precipitation. If central venous catheter occlusion does occur due to precipitation of calcium phosphate crystals, then HCl-heparin irrigation is a safe and effective method for salvaging such catheters.

A method for toxicological evaluation of biomaterials based on colony formation of V79 cells.

This report describes a method for cytotoxicity screening of biomaterials based on colony formation of V79 cells. For this test, two metals (titanium and nickel), two ceramics (alumina ceramic and tricalcium phosphate), and two types of polymeric material [high density polyethylene (HDP) and polyvinylchloride (PVC)] were used. Each metal and ceramic was cast into a disk and semidisk 49 mm in diameter and 1 to 2 mm thick. The HDP was molded into a petri dish and PVC was used as a thin film. The materials were sterilized by heating or with ethylene oxide and placed in plastic petri dishes, after which 8 ml cell suspension containing 100 cells were added to each dish. After 1 week, the colonies formed on the materials were fixed, stained, and then the number of colonies was counted. Titanium, alumina ceramic, and HDP showed no differences from the controls in terms of colonies. On the disks and the semidisks of nickel and tricalcium phosphate and on the thin disks of PVC, however, no colonies were detected. The V79 cells used in this experiment showed a rapid and logarithmically stable growth curve and such a high rate of colony formation as to form visible noticeable colonies, and were therefore suitable cells for screening test the cytotoxicity of biomaterials. Unlike other previously reported methods of in vitro cytotoxicity testing, this method permits assay of colonies formed from a single cell after proliferation directly on the materials. Moreover, the test with semidisks permits simple screening to assess the cytotoxicity is caused by either the chemical substances or the physical properties of the materials.(ABSTRACT TRUNCATED AT 250 WORDS)