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At least 19 recordsLinked to original sources

Kinetics of cortical bone demineralization: controlled demineralization--a new method for modifying cortical bone allografts.

We investigated the kinetics of hydrochloric acid demineralization of human cortical bone with the objective of developing a method of controlled demineralization for structural bone allografts. It is known that the demineralization of cortical bone is a diffusion rate limited process with a sharp advancing reaction front. The demineralization kinetics of human cortical bone, described as the advance of the reaction front versus immersion time, were determined by measuring extraction of bone mineral in both planar and cylindrical geometries. Mathematical models based on diffusional mass transfer were developed to predict this process. The experimental data fit well with the behavior predicted by the model. The model for planar geometry is applicable to controlled demineralization of cortical bone allografts of irregular shapes such as cortical struts. The model for cylindrical geometry is appropriate when curved surfaces are involved such as in diaphyseal bone allografts. This method of demineralization has direct application to clinical modification of cortical bone allografts to potentially enhance their osteoinductive properties.

Adult↗

Staining of demineralized cartilage. II. Quantitation of articular cartilage proteoglycan after fixation and rapid demineralization.

Safranin O in the orthochromatic form stains articular cartilage proteoglycan quantitatively in histological sections of demineralized cartilage. This was shown by scanning microdensitometry of stained sections of undemineralized and demineralized articular cartilage and by biochemical analysis of 35S labelled cartilage subjected to demineralization. In contrast, Alcian Blue staining is affected by unknown factors other than simply the amount of proteoglycan present. Alcoholic formalin fixes articular cartilage proteoglycan more successfully than formol Zenker for subsequent rapid demineralization. Alcoholic formalin does not preserve cellular appearance as well as formol Zenker. Staining of articular cartilage with PAS appears unaffected by demineralization.

Aged↗

Effect(s) of the demineralization process on the osteoinductivity of demineralized bone matrix.

The relationships between residual calcium levels and particle size of ground demineralized bone matrix and its osteoinductive potential were investigated using in vitro and in vivo assays. The effects of variable residual calcium levels, variable particle sizes, and donor age and gender were studied using a tissue culture-based bioassay (in vitro) as well as an athymic mouse (in vivo) bioassay. The osteoinductive potential of the bone-derived biomaterial was assessed by measuring the degree of new bone formation (change in percent calcium content after 4 weeks of implantation) in the in vivo assay and levels of alkaline phosphatase activity associated with cultures of human periosteal cells (HPO cells) in the in vitro assay, respectively. Slightly demineralized bone matrix and overly demineralized bone matrix possessed a degree of osteoinductive potential whereas bone demineralized to levels of approximately 2% residual calcium provided for maximum osteoinductive potential in both assay systems. The osteoinductive potential of ground demineralized bone varied relative to the particle size such that DBM particles ranging from 500 to 710 microns provided for the highest level of calcium deposition (increase of 8.1 weight percent calcium) after 4 weeks of implantation in muscle pouches of an athymic mouse, whereas explanted particles less than 250 microns showed the lowest level of calcium deposition (increase of only 2.8 weight percent calcium). In the donor age and gender study, DBM from different donors were divided into 5 age groups for both female and male donor derived bone: less than 20, 21 to 30, 31 to 40, 41 to 50, and 51 to 60 year old age groups. This study indicated that DBM from female donors in the 31 to 40 years old age group and male donors in the 41 to 50 year age group possess the highest osteoinductive potential, whereas DBM derived from donor bone from both female and male donors in the 51 to 60 year age group presented the lowest osteoinductive potential. DBM derived from male and female donors did not in general show significant differences in osteoinductive potential.

Adult↗

Staining of demineralized cartilage. I. Alcoholic versus aqueous demineralization at neutral and acidic pH.

Demineralization of cartilage with alcoholic EDTA provides cartilage staining that is no better, as measured by scanning microdensitometry, than that of adequately fixed specimens demineralized with aqueous EDTA. Aqueous EDTA is a faster demineralizing agent than alcoholic EDTA. Certain fixatives can preserve maximal proteoglycan staining in articular cartilage even with subsequent rapid demineralization in formate buffer at pH 3.3. Although alcoholic formalin fixation provided optimum quantitative cartilage staining, cetylpyridinium chloride (CPC) in aqueous buffered formalin improved cellular detail, but CPC partially suppressed matrix staining.

Animals↗

The inhibition of demineralization of human enamel after fluoride varnish application as a function of the fluoride content. An in vitro study under constant composition demineralizing conditions.

The inhibiting effect of a 24 hours application of a fluoridated varnish with various fluoride contents on demineralization of human sound enamel was evaluated in vitro. The varnishes used had the same polyurethane base (Fluor Protector) and contained 0.7; 0.1; 0.05 and 0 wt% fluoride, resp. A constant composition technique was used to demineralize varnished and non-varnished specimens at a pH of 5 for periods upto 2 weeks. Microhardness measurements were carried out after several time intervals to follow mineral loss in time longitudinally. At the end of each experimental run microradiography was carried out to investigate 1) lesion type, 2) lesion depth and 3) mineral loss. It is shown in this study that the fluoride releasing varnishes applied on the enamel for 24 hours can inhibit demineralization completely. No demineralization inhibition with the 0% fluoride varnish application was observed.

Decalcification Technique↗

The effect of triclosan toothpaste on enamel demineralization in a bacterial demineralization model.

Triclosan has been incorporated into toothpaste to enhance inhibitory effects on bacterial metabolism in dental plaque. Many studies have confirmed these effects by showing a reduction of accumulation of dental plaque, gingivitis and calculus. However, there is no evidence for triclosan having an inhibitory effect on the dental plaque-induced demineralization of the dental hard tissues. Therefore, the effect of 0.3% triclosan added to non-fluoride and fluoride toothpaste was tested in an in vitro model, in which bovine enamel specimens were to be demineralized by acids produced in overlaying Streptococcus mutans suspensions. In a first set of experiments the toothpastes were added to the S. mutans suspensions at 1:100, 1:1000 and 1:10,000 (w/v) dilutions. After 22 h incubation at 37 degrees C the suspensions were removed and assessed for calcium and lactate content, and pH. In this set of experiments, triclosan had no additive protective effect to the non-fluoride or fluoride toothpaste. In a second set of experiments, the enamel specimens were immersed daily for 3 min in 30% (w/v) slurries of the toothpastes before the 22 h incubation with the S. mutans suspensions. Under these conditions, triclosan showed an additional protective effect compared with non-fluoride toothpaste at a low concentration of S. mutans cells (0.07 mg cells dry weight per 600 microL suspension). It is concluded that the enamel surface may act as a reservoir for triclosan, which may protect the enamel surface against a mild acid attack. In combination with fluoride, however, as in toothpaste, triclosan has no additional protective effect against demineralization.

Animals↗

A comparison of leakage of filling materials in demineralized and non-demineralized resected root ends under vacuum and non-vacuum conditions.

A study was conducted to compare the marginal apical leakage of root-end filling materials under vacuum and non-vacuum conditions, and to assess the effect of demineralization by application of citric acid to the root ends on the apical marginal leakage of root end filling materials. The root canals of 148 extracted human canines and premolars were handfiled and sealed with gutta-percha and Roth's 801 sealer using cold lateral condensation. The teeth were divided into 8 groups that received retrofilling materials. The materials tested were amalgam, IRM and gutta-percha/sealer with and without successive application of citric acid. Half of the specimens were placed under vacuum conditions in methylene blue dye, and the other half were placed in the dye for the same time period without exhausting air from the flask. The teeth were split longitudinally and the extent of dye penetration was determined with a stereomicroscope and eyepiece micrometer. The most evident difference in dye penetration was found between the positive control groups. Under vacuum conditions, the positive controls showed complete penetration of the unobturated canal system with dye. With passive immersion only partial penetration of the unobturated canal system occurred. Application of citric acid to the root end did not adversely affect the seal of amalgam, IRM or gutta-percha/sealer.

Acid Etching, Dental↗

In vitro demineralization by strains of Actinomyces viscosus and Streptococcus sobrinus of sound and demineralized root surfaces.

Root sections were inoculated with one of two strains of Actinomyces viscosus or a strain of Streptococcus sobrinus and then incubated for 9 days in Trypticase soy broth (TSB) containing 0.25% glucose or TSB supplemented with 0.25% sucrose (TSB+S). Lesion progression was measured from microradiographs. One section from each group was examined with a transmission electron microscope. Lesion progression was associated with a fall in the pH of the medium. However, lesion progression was not correlated with the mean 48-hour pH of the medium. In both TSB and TSB + S, lesion progression with A. viscosus was significantly greater than in the S. sobrinus group. However, the mean 48-hour pH value in TSB + S was lower than that in the S. sobrinus group. Plaque formation in TSB was, subjectively, least in the S. sobrinus group. Examination of transmission electron micrographs revealed bacteria penetrating the surfaces of the sections and extending into the lesion in the A. viscosus groups but not in the S. sobrinus group. This in vitro bacterial plaque model of root caries may be suitable for investigations of the bacterial etiology of root-surface caries and the virulence factors associated with pathogenicity.

Actinomyces viscosus↗

Platelet-rich plasma inhibits demineralized bone matrix-induced bone formation in nude mice.

BACKGROUND: It is unclear whether platelet-rich plasma is a clinically effective adjunct to osteoinductive agents such as demineralized bone matrix. It contains platelet-derived growth factor (PDGF), which decreases osteoinduction by human demineralized bone matrix in nude-mouse muscle, suggesting that platelet-rich plasma may also have a negative impact. This study tested the hypothesis that platelet-rich plasma reduces demineralized bone matrix-induced bone formation and that this effect varies with donor-dependent differences in platelet-rich plasma and demineralized bone matrix. METHODS: Human platelet-rich plasma was prepared from blood from six men (average age [and standard error of the mean], 29.2 +/- 2.4 years). Platelet numbers were determined, and growth factors were quantified before and after platelet activation. Human demineralized bone matrix from two donors (demineralized bone matrix-1 and demineralized bone matrix-2) was mixed with activated platelet-rich plasma and was implanted bilaterally in the gastrocnemius muscle in eighty male nude mice (eight implants per variable). Fifty-six days after implantation, the hindlimb calf muscles were harvested for histological analysis. Osteoinduction was evaluated with use of a qualitative score and morphometric measurements of ossicle size, new bone formation, and residual demineralized bone matrix. RESULTS: Compared with platelet-poor plasma, platelet-rich plasma preparations exhibited a fourfold increase in the platelet count, a fifteenfold increase in the amount of transforming growth factor-beta, a sixfold increase in the amount of PDGF-BB, a fivefold increase in the amount of PDGF-AA, and a twofold increase in the amount of PDGF-AB. Demineralized bone matrix-1 was more osteoinductive than demineralized bone matrix-2, as determined on the basis of a greater ossicle area. The effect of platelet-rich plasma was either neutral or inhibitory depending on the demineralized bone matrix batch. When used with demineralized bone matrix-1, platelet-rich plasma did not alter the qualitative score or overall ossicle size, but it decreased the new bone area. When used with demineralized bone matrix-2, platelet-rich plasma reduced the qualitative score, ossicle area, and new bone area and increased the amount of residual demineralized bone matrix. The effects on osteoinduction also varied with the donor of the platelet-rich plasma. CONCLUSIONS: Platelet-rich plasma decreased the osteoinductivity of demineralized bone matrix implanted in immunocom-promised mice, and the activities of both demineralized bone matrix and platelet-rich plasma were donor-dependent.

Adult↗

Experimental posterolateral lumbar spinal fusion with a demineralized bone matrix gel.

STUDY DESIGN: A controlled rabbit model of lumbar posterolateral intertransverse process arthrodesis was used to evaluate a bone graft substitute. OBJECTIVE: To determine the efficacy of demineralized bone matrix gel as an autograft extender, using different ratios of demineralized bone matrix to autograft and to determine the efficacy of demineralized bone matrix as an autogenous bone graft enhancer by adding it to the usual quantity of autograft. SUMMARY OF BACKGROUND DATA: Autogenous bone is considered the most effective bone graft material for posterolateral lumbar arthrodesis, yet nonunions occur in up to 30% of patients. In addition, donor site complications may occur in 25-30% of patients. This has prompted the search for and investigation of bone graft extenders, enhancers, and substitutes. Commercially available demineralized bone matrix gel is one possible graft extender and enhancer, which, unlike mineralized allografts, has osteoinductive properties. Although the gel is in common use, the efficacy of demineralized bone matrix when used for posterolateral spine arthrodesis has not been examined in prospective clinical studies. Furthermore, no known animal studies have tested demineralized bone matrix gel in a posterolateral arthrodesis model. METHODS: Forty-seven New Zealand white rabbits underwent bilateral posterolateral spine arthrodesis at L5-L6 using autogenous iliac crest bone graft alone or in combination with demineralized bone matrix. Four groups were formed on the basis of the ratio of autograft to demineralized bone matrix: autograft alone (3 mL), 100:0 group; autograft (3 mL) and demineralized bone matrix (1.5 mL), 100:50 group; autograft (1.5 mL) and demineralized bone matrix (1.5 mL), 50:50 group; and autograft (0.75 mL) and demineralized bone matrix (2.25 mL), 25:75 group. Rabbits were killed 6 weeks after surgery. Inspection, manual palpation, radiographic film, and histologic evaluation were used to assess fusion. RESULTS: All groups had similar fusion rates (66-73%) based on manual palpation. Rabbits implanted with demineralized bone matrix had more mature fusion masses, evidenced by the greater trabecular bone formation seen on radiographic film and histologic study. CONCLUSIONS: Demineralized bone matrix was effective as a graft extender when used in up to a 3:1 ratio with autograft in a rabbit posterolateral spine fusion model. When less than the standard volume of autograft was used, the addition of demineralized bone matrix gel lead to fusion success rates comparable to those of the standard amount of autograft alone. However, demineralized bone matrix did not increase the frequency of successful fusion when added to the standard amount of autograft.

Animals↗

Osteoinduction of human demineralized bone: characterization in a rat model.

The increasing clinical use of human demineralized bone matrix has brought about the desire to understand better the osteoinductivity of these graft materials. The rat heterotopic model has been used successfully to show the sequence of events involved in the endochondral ossification process resulting in osteoinduction. In this study, the osteoinductive potential of human demineralized bone powder was assessed, using immune compromised rats (athymic rnu/rnu) to avoid problems associated with cross species incompatibilities. Implants were placed in subcutaneous or intermuscular sites. This model is characterized to provide a basis for routinely determining the performance of human demineralized bone powder. Demineralized bone powder was prepared from rat and human cortical bone according to a strict protocol. The lack of response to guanidine HCl extracted (noninductive) demineralized bone showed the selectivity of the assay. The same lots of human and rat demineralized bone were tested in sequential experiments during a 1-year period. These results showed reproducible induction of the demineralized bone powder between experiments. Combining demineralized bone with the guanidine HCl extracted demineralized bone in varying ratios tested the sensitivity of the assay. These results showed an increase in bone formation with increasing quantities of active demineralized bone and established the ability of the bioassay to differentiate between the various levels of active (osteoinductive) demineralized bone powder. With this model, consistent performance of demineralized bone powder processed by well controlled methods was seen.

Animals↗