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PubMed1989

[Bonding of MMA-TBB resin to bovine tooth coated by poly (methyl methacrylate-co-p-styrene sulfonic acid)].

The objective of this study was to prepare a new type water soluble bonding agent, methyl methacrylate (MMA)-p-styrene sulfonic acid copolymer (MS), and to investigate the effect of MS on bonding between resins and tooth substrates. MS is cross-linked with Ca2+ supplied by hydroxyapatite in a smeared layer on ground enamel and dentin and sticks to their surface. Samples were prepared by bonding an acrylic rod with MMA-TBB resin to ground enamel and dentin coated with an aqueous mixture of FeCl3 and 10 wt% MS. After immersed in water for 24 hrs, the tensile bond strength was measured. The bond strengths to both enamel and dentin were higher than 11 MPa and cohesive failure of cured MMA-TBB resin was observed in every case. This suggested that MS could adhere to tooth substrates with a new bonding mechanism different from the previously reported mechanism of the monomer interpenetration and polymerization.

Adhesives↗
PubMed2001

A fluoride-releasing composite for dental applications.

OBJECTIVES: The aim of this work was to attempt formulation of a dental composite, for investigational purposes, which will release fluoride at a rate comparable to glass ionomer materials, while retaining physical properties adequate for potential use in a number of dental applications. METHODS: An organic fluoride material, tetrabutylammonium tetrafluoroborate (TBATFB), was incorporated into a hydrophilic monomer system made of 2,2-bis[4-(2-hydroxy-3-methacroyloxypropoxy)phenyl]-propane (BisGMA) and 2-hydroxyethyl methacrylate (HEMA). This fluoride-containing monomer system was filled with silane treated quartz to 81 wt%. The resultant material was tested for fluoride release, physical strength properties and bond strength to enamel, each in comparison to control materials. RESULTS: Cumulative fluoride release from the experimental composite was approximately linear over time and comparable to glass ionomer materials. The long term rate of release exceeded several of the glass ionomer materials tested. Diametral tensile strength was comparable to resin-modified glass ionomer (RMGI) materials. Bond strength to etched enamel was statistically equivalent to a commercially available hybrid composite control. SIGNIFICANCE: The experimental resin-based composite material could potentially be used as an alternative to glass ionomer materials in non-stress bearing restorative applications and for orthodontic bracket bonding, where high fluoride release is desirable.

Acid Etching, Dental↗
PubMed1998

Development of glass-ionomer cement systems.

In the 1960s the idea of positive physico-chemical adhesion with tooth substance resulted in the invention of polyacrylic acid-based cements, first the zinc polycarboxylate and, subsequently, the glass-ionomer cements. These materials were shown to undergo specific adhesion with hydroxyapatite and proved to have properties satisfactory for a variety of clinical applications. The key properties of the glass-ionomer cements--fluoride release over a prolonged period and specific adhesion to enamel and dentine coupled with aesthetic qualities are related to their characteristics as aqueous polyelectrolyte systems. In order to improve toughness, speed of setting and resistance to dehydration, hybrid materials in which some of the water content of the glass-ionomer system was replaced by water-soluble polymers or monomer systems capable of ambient polymerization were formulated in the late 1980s. These materials, which have been termed resin-modified glass-ionomer cements, involve, ideally, the formation of an interpenetrating polymer network combining the acid-base cross-linking reaction of the metal ion-polyacid with the cross-linking polymerization of the monomer system or additive action of the polymers. In the predominantly resin materials there is little polyelectrolyte character and it is controversial whether such materials should be categorized as glass-ionomer cement systems. The specific advantages of these materials over traditional glass-ionomer systems and over composite restorative systems remain to be fully documented. Studies of adsorption to hydroxyapatite of typical monomers using X-ray photoelectron spectroscopy (XPS) and time-of-flight secondary ion mass spectrometry (TOF SIMS) indicate that resistance to water displacement decreases as hydrophobicity increases.

Biocompatible Materials↗
PubMed1997

Formulation of enamel matrix derivative for surface coating. Kinetics and cell colonization.

Enamel Matrix Derivative (EMD) contains a protein complex belonging to the amelogenin family. Enamel matrix as well as EMD have been found to promote periodontal regeneration when applied onto denuded root surfaces in dehiscence models. In the present studies it is shown that propylene glycol alginate (PGA) is a suitable vehicle for EMD for its local application. EMD can be dissolved in PGA at an acidic pH, resulting in a highly viscous solution. At neutral pH and body temperature the viscosity decreases and EMD precipitates. Multilayers of EMD on mineral or protein surfaces have been analysed using ellipsometry, total internal reflection fluorescence (TIRF) and biospecific interaction analysis (BIA). The studies show that EMD adsorbs both to hydroxyapatite and collagen and to denuded dental roots. It forms insoluble spherical complexes, and detectable amounts remain at the site of application on the root surface for two weeks, as shown with radiolabelled protein in rats and pigs. Scanning electron micrograph (SEM) studies on monkey teeth further indicate that EMD in PGA may promote repopulation of fibroblast-like cells during the first weeks after application.

Adsorption↗
PubMed1983

The dynamics of formation of a collagen-phosphophoryn conjugate in relation to the passage of the mineralization front in rat incisor dentin.

Dentin and predentin matrices contain Type I collagen and phosphophoryns as major constituents. A collagen-phosphophoryn conjugate is also present in small amounts. This conjugate has been implicated in the deposition of mineral. Its formation has been followed in rat incisors. Rats were labeled for varied time intervals with [3H]proline, followed by a 2-h pulse of [3H] serine. The soluble alpha- and beta-phosphophoryns were extracted under conditions minimizing degradation. The tooth residue was CNBr-treated and the collagen CNBr peptides alpha 1(I)CB7 and alpha 1(I)CB8 were collected along with the solubilized conjugate fraction. Each component was purified and the specific activities in [3H] proline, [3H]hydroxyproline, [3H]serine, and [3H]phosphoserine were determined. The collagen and alpha-phosphophoryn accumulated proline label linearly at the same rate over the entire period of labeling. Entry of [3H]proline into the conjugate fraction was delayed by approximately 9-10 h and then the label accumulated also linearly at the same rate. [3H]Serine was present at a different but constant level in each fraction; the conjugate had the lowest activity. These data indicate an extracellular formation of the conjugate at the mineralization front from precursors which followed different secretory pathways.

Amino Acids↗
PubMed1979

Serially etched shark enameloid observed by incident light microscopy.

Longitudinal and transverse tooth sections of Isurus oxyrinchus were serially etched in 2.6% nitric acid. The changing optical properties of the etched surfaces were observed during the serial etchings, and the descent of the enameloid surfaces was measured. Shark enameloid seems to be less effectively etched by acid than human enamel; this difference may be due to differences in solubility between fluorapatite and hydroxyapatite. Most of the information regarding the structure of the enameloid was gained during the first five of ten etchings. The reflection of light from the surface was influenced by the orientation of the crystallites, longitudinally sectioned crystallites reflecting the light better than transversely sectioned crystallites. The dentinal extensions were continuous with and of the same structure as the underlying dentine. The radial fibers originated from the dentinal extensions, and they both contained organic material and were accompanied by crystallites. When the specimens were imbibed with water the distinctness of the dentinal extensions and radial fibers was improved.

Acid Etching, Dental↗
PubMed2005

Inhibitory effect of CGRP on osteoclast formation by mouse bone marrow cells treated with isoproterenol.

The present study was designed to elucidate the mode of action of isoproterenol (Isp; adrenergic beta-agonist) and to characterize the effect of the calcitonin gene-related peptide (CGRP; sensory neuropeptide) on osteoclast formation induced by Isp in a mouse bone marrow culture system. Treatment of mouse bone marrow cells with Isp generated tartrate-resistant acid phosphatase (TRAP)-positive multinuclear cells (MNCs) capable of excavating resorptive pits on dentine slices, and caused an increase in receptor activator of NF-kappaB ligand (RANKL) and a decrease in osteoprotegerin (OPG) production by the marrow cells. The osteoclast formation was significantly inhibited by OPG, suggesting the involvement of the RANKL-RANK system. CGRP inhibited the osteoclast formation caused by Isp or soluble RANKL (s-RANKL) but had no influence on RANKL or OPG production by the bone marrow cells treated with Isp, suggesting that CGRP inhibited the osteoclast formation by interfering with the action of RANKL produced by the Isp-treated bone marrow cells without affecting RANKL or OPG production. This in vitro data suggest the physiological interaction of sympathetic and sensory nerves in osteoclastogenesis in vivo.

Adrenergic beta-Agonists↗
PubMed1999

Altered phosphorylation of rat dentine phosphoproteins by fluoride in vivo.

Dentine phosphoproteins have been proposed to have an important role in mineralization. This study focused on the influence of fluoride on the biochemical composition of dentine phosphoproteins and attempts to relate changes to the altered mineralization witnessed during fluorosis. Wistar rats were rendered fluorotic by the administration of 20 ppm sodium fluoride in their drinking water ad libitum, a nonfluorotic group received double-distilled, deionized water only. After 17 weeks, the teeth showed signs of fluorosis. The incisors were removed, split longitudinally, and the pulps were removed. Teeth were powdered and demineralized in 10% EDTA with protease inhibitors, after which the organic matrix was extracted with 4 M guanidinium chloride. Phosphoproteins were selectively precipitated from the soluble extract by the addition of 1.0 M calcium chloride and further purified by anion exchange chromatography. SDS-PAGE revealed two protein bands with molecular weights of 130 kDa and 66 kDa in the nonfluorotic fraction and 116 kDa and 66 kDa in the fluorotic fraction. Western blotting analysis identified the 66 kDa band as alpha2-HS glycoprotein which co-precipitated with phosphoproteins. Electroelution of the protein bands was performed with subsequent biochemical analyses. Phosphate content was determined for each protein band and was detectable in the 116 kDa and 130 kDa bands from the fluorotic and nonfluorotic samples, respectively, with a decreased level noted in the 116 kDa band. The presence of phosphate and the amino acid analysis of these bands suggested their identity to be dentine phosphoproteins. No changes in the ratio of amino acids was detected in fluorotic samples. The fluoride-induced alterations to the biochemical structure of dentine phosphoproteins would appear to influence the phosphorylation of these macromolecules only, possibly affecting posttranslational events. Such alterations may play a role in disrupting the patterns of mineralization seen during fluorosis.

Amino Acids↗
PubMed2004

Interleukin (IL) 18 stimulates osteoclast formation through synovial T cells in rheumatoid arthritis: comparison with IL1 beta and tumour necrosis factor alpha.

OBJECTIVE: To determine whether IL18 has any indirect effects on osteoclastogenesis mediated by T cells in RA synovium, and compare its effects with those of IL1 beta and TNF alpha. METHODS: Resting T cells were isolated from peripheral blood of healthy donors, and stimulated with 2 microg/ml phytohaemagglutinin (PHA) and 0.5 ng/ml IL2 for 24 hours. Synovial T cells were isolated from RA synovial tissue. The levels of soluble receptor activator of the NF-kappa B ligand (RANKL), osteoprotegerin (OPG), IFN gamma, M-CSF, and GM-CSF were determined by ELISA. Membrane bound RANKL expression was analysed by flow cytometry. Commercially available human osteoclast precursors were cocultured with T cells to induce osteoclast formation, which was determined with tartrate resistant acid phosphatase staining and pit formation assay. RESULTS: In PHA prestimulated T cells or RA synovial T cells, IL18, IL1 beta, or TNFalpha increased soluble RANKL production and membrane bound RANKL expression in a dose dependent manner. IL18, IL1 beta, and TNF alpha did not induce M-CSF, GM-CSF, IFN gamma, or OPG production in PHA prestimulated T cells or RA synovial T cells. IL18 increased the number of osteoclasts and bone resorption area on dentine slices in the coculture of human osteoclast precursors with PHA prestimulated T cells or RA synovial T cells; its ability was equivalent to that of IL1 beta, but less potent than that of TNF alpha. In the coculture system, OPG completely blocked osteoclast induction by IL18 or IL1 beta, and greatly inhibited induction by TNF alpha. CONCLUSION: IL18, IL1 beta, or TNF alpha can indirectly stimulate osteoclast formation through up regulation of RANKL production from T cells in RA synovitis; IL18 is as effective as IL1 beta, but less potent than TNF alpha.

Analysis of Variance↗
PubMed2000

Effect of osteoprotegerin and osteoprotegerin ligand on osteoclast formation by arthroplasty membrane derived macrophages.

OBJECTIVE: Osteoprotegerin ligand (OPGL) is a newly discovered molecule, which is expressed by osteoblasts/bone stromal cells. This ligand and M-CSF are now known to be essential for osteoclast differentiation from marrow and circulating precursors. This study examined whether OPGL and its soluble receptor osteoprotegerin (OPG), influenced osteoclast formation from human arthroplasty derived macrophages, to determine if the effects of OPGL and OPG on these cells could contribute to the osteolysis of aseptic loosening. METHODS: OPGL (+/- dexamethasone/M-CSF) was added to cultures of macrophages isolated from the pseudomembrane of loosened hip arthroplasties incubated on glass coverslips and dentine slices. OPG was added to cocultures of arthroplasty derived macrophages and UMR106 osteoblast-like cells. Osteoclast differentiation in long term cultures was assessed by expression of macrophage (CD14) and osteoclast markers (tartrate resistant acid phosphatase (TRAP), vitronectin receptor (VNR) and lacunar resorption). RESULTS: In the absence of osteoblastic cells, the addition of OPGL alone was sufficient to induce differentiation of macrophages (CD14(+), TRAP(-), VNR(-)) into TRAP(+) and VNR(+) multinucleated cells, capable of extensive lacunar resorption. OPG was found to inhibit osteoclast formation by arthroplasty macrophages in a dose dependent manner. OPG (100 ng/ml) more than halved the formation of TRAP(+) and VNR(+) cells and the extent of lacunar resorption in co-cultures of UMR106 cells and arthroplasty macrophages. CONCLUSIONS: This study has shown that macrophages, isolated from the pseudomembrane surrounding loose arthroplasty components, are capable of differentiating into osteoclastic bone resorbing cells and that OPGL is required for this to occur. OPG inhibits this process, most probably by interrupting the cell-cell interaction between osteoblasts and mononuclear phagocyte osteoclast precursors present in the pseudomembrane.

Aged↗
PubMed2001

Tumor necrosis factor-alpha cooperates with receptor activator of nuclear factor kappaB ligand in generation of osteoclasts in stromal cell-depleted rat bone marrow cell culture.

A member of the tumor necrosis factor (TNF) family, receptor activator of nuclear factor kappaB ligand (RANKL; also known as ODF, OPGL, and TRANCE), plays critical roles in osteoclast differentiation and activation in the presence of macrophage colony-stimulating factor (M-CSF). Recently, TNF-alpha has also been shown to induce the formation of multinucleated osteoclast-like cells (MNCs) in the presence of M-CSF from mouse macrophages. We demonstrated that mononuclear preosteoclast-like cells (POCs) were formed in the presence of conditioned medium of osteoblastic cells in a rat bone marrow culture depleted of stromal cells. Using this culture system, in this study we examined whether TNF-alpha affects differentiation into POCs from hematopoietic progenitor cells. Human TNF-alpha (hTNF-alpha) markedly stimulated the formation of POCs. Moreover, a concentration as low as 0.005 ng/mL of hTNF-alpha increased the level of mRNA for calcitonin receptor (CTR) and cathepsin-K of POCs. The POCs induced by hTNF-alpha formed MNCs, which showed dentine-resorbing activity after coculture with primary osteoblasts. Stimulation was observed after 24 h of treatment with hTNF-alpha only on day 1 or day 2 of the culture. After 24 h of hTNF-alpha treatment, expression of the receptor activator of nuclear factor kappaB (RANK) mRNA was markedly increased. The addition of soluble RANKL (sRANKL) to the preformed POCs efficiently induced MNCs. Interestingly, treatment of bone marrow cells with hTNF-alpha and sRANKL synergistically augmented the formation of MNCs. This formation was abolished by the addition of human osteoprotegerin (hOPG). These results suggest that cooperation of TNF-alpha and RANKL is important for osteoclastogenesis.

Animals↗
PubMed1990

[Biochemical study on the application of alpha-cyanoacrylate instant adhesives in dentistry].

The author biochemically evaluated the possible application of 4 types of alpha-cyanoacrylate adhesive agents (methyl, ethyl, isopropyl and isobutyl) in dentistry. A semiconductor temperature sensor was used to measure alpha-cyanoacrylate polymerization-temperature maximum heat, and an oscilloscope was employed to measure heat value. Amounts of formaldehyde eluted from the soluble component of alpha-cyanoacrylate was measured colorimetrically. In addition, to evaluate the effects of alpha-cyanoacrylate on pulp tissue, the soluble component of alpha-cyanoacrylate polymer was added to mouse macrophages. Changes in the numbers and morphology of macrophages and their glucose consumption in the supernatant medium were studied and compared with corresponding data obtained when formaldehyde alone was used. (1) During the polymerization of alpha-cyanoacrylate, when a glass fiber disc filter was used, the temperature increased by 19.1 degrees C in 1 microliter with methyl and by 8.6 degrees C with isobutyl. Even with the same alkyl, however, polymerization heat differed depending on the structure and constitution of the adhesive material and varied widely according to experimental conditions. (2) A mean maximum temperature of 1.7 degrees C in the dentin floor during polymerization of 10 microliters of alpha-cyanoacrylate monomer with methyl suggests no thermal injury to the pulp. (3) Formaldehyde was detected in all 4 adhesives when alpha-cyanoacrylate was dropped into distilled water for polymerization. Since, with each alkyl, the amount of eluted formaldehyde reached a maximum after 1 day of immersion, continuous elution of formaldehyde by hydrolysis was negligible. (4) A greater amount of formaldehyde was eluted from the polymer with methyl and ethyl, which have less Cs, than from the polymer with isopropyl and isobutyl, which have more Cs. These findings suggest that alkyl type affects the amount of eluted formaldehyde. (5) The soluble component of the alpha-cyanoacrylate polymer, either methyl or ethyl, had no effect on the number or morphology of mouse macrophages, as compared with corresponding data obtained when sterilized distilled water was used as a control. Isopropyl and isobutyl, however, decreased cell count and inhibited the extension of the sell soma. (6) The effects of isopropyl and isobutyl on the numbers and morphology of macrophages were similar to those of formaldehyde used alone at a concentration of 8 ppm or more. These findings suggest that, under the present experimental conditions, the above-mentioned changes in macrophages are the effects of about 8 ppm of formaldehyde. (7) The rate of glucose consumption by macrophages with methyl or ethyl was as high as that with the control.(ABSTRACT TRUNCATED AT 400 WORDS)

Adhesives↗
PubMed1991

Isolation of a murine osteoclast colony-stimulating factor.

Cultures of a cell line derived from a murine mammary carcinoma that induces hypercalcemia were examined for soluble products that could induce osteoclasts to differentiate from murine bone marrow cells. The serum-free culture supernatant of this cell line stimulated growth of colonies from bone marrow cells that exhibited tartrate-resistant acid phosphatase (TRAPase) activity. These TRAPase-positive cells demonstrated essential features of osteoclasts when cultured with mineralized bone or dentin. The culture period required for colony development and the frequency of colony-forming cells indicated that relatively primitive marrow progenitors were stimulated by a tumor-derived factor(s) to form immature osteoclasts. Other colony-stimulating factors (CSFs), including granulocyte CSF, macrophage CSF, granulocyte-macrophage CSF and interleukin 3, were ruled out as the source of the activity produced by the tumor cells. The biological activity was successfully purified by gel filtration chromatography and reverse-phase HPLC. By SDS/PAGE, the activity was traced to a protein of approximately 17 kDa. Functional and biochemical studies of the purified factor suggest that it is distinct from any known CSF of myeloid cells. This protein appears to be a CSF for the osteoclast lineage, osteoclast CSF (O-CSF).

Acid Phosphatase↗
PubMed1994

Three-dimensional reconstruction of microleakage pattern using a sequential grinding technique.

Dye penetration tests are very commonly used to detect the absence of a fluid seal at the tooth-restoration interface. Airlocks in the marginal gap, leaching of water-soluble tracers during processing, and the failure of only a few sections to allow interpretation of the full pattern, limit these tests to low reproducibility and precision. The purpose of this present study was to generate high-resolution three-dimensional images of waterfast tracer patterns. Cylindrical class V (3 mm diameter, 2 mm deep) dentine-bonded resin composite restorations in buccal coronal dentine were thermally cycled (1000 x, 8 degrees C, 55 degrees C, 30 s dwell at each temperature) and then silver stained using an initial vacuum (100 mmHg pressure). Each restoration was sequentially abraded from the free surface on wet 180 grit silicon carbide paper, producing up to 30 parallel surfaces at approximately 0.15 mm separation through the restoration down to the pulp. Images of the ground surfaces were captured, and assembled by a computer image analyser program to give a three-dimensional model of the tracer pattern. The maximum depths of tracer penetration below the reference surfaces were 3.00 mm, 2.09 mm, 3.16 mm and > 2.29 mm for the four specimens. Projections of the models were viewed from several directions with sections in various locations to allow investigation of the full tracer pattern. This method allows the creation of high-resolution three-dimensional tracer patterns.

Composite Resins↗
PubMed1990

High resolution scanning electron microscopy of the subplasmalemmal cytoskeleton in human odontoblasts.

The subplasmalemmal cytoskeleton in human odontoblasts was studied by high resolution scanning electron microscopy. The odontoblast layer was isolated and exposed to formaldehyde, glutaraldehyde, and OsO4 for some specimens, while the membraneous structures and soluble proteins in the dental tissue were removed by Zenker's solution and 1% OsO4 for other specimens, without further fixation of the remaining components. The cytoskeletal elements comprised a dense network of interlacing filaments of different diameters in the cell body. Most cytoskeletal elements were parallel to the axis of the cell processes situated inside the dentinal tubules. The appearance and orientation of the investigated subplasmalemmal cytoskeletal elements was unaffected by the choice of method. Both methods confirm the presence of a subplasmalemmal cytoskeleton in human odontoblasts.

Actin Cytoskeleton↗
PubMed2000

Transforming growth factor-beta isoform expression in mature human healthy and carious molar teeth.

Transforming growth factor (TGF)-beta isoforms have been implicated in cellular signalling during tooth development and repair, but little is known of their cellular localisation or distribution within the dental tissues in the mature tooth. This study investigated the presence of TGF-beta1, beta2 and beta3 isoforms in tissues of sound and carious human molar teeth, to understand better the expression of TGF-betas during health and disease. In healthy tissues, odontoblasts, cells of the cell rich layer, pulpal fibroblasts and endothelial cells were stained to varying degrees for all isoforms, with TGF-beta3 showing the greatest intensity and TGF-beta1 the weakest intensity. Similar patterns of staining were observed in carious teeth; however, TGF-beta1 showed significantly increased staining intensity within odontoblasts and pulpal cells of carious teeth (p < 0.001). Biochemical analysis showed greater amounts of TGF-beta1 in tertiary dentine than in primary dentine samples. The expression of TGF-betas in odontoblasts and the increased presence of TGF-beta1 in tertiary dentine suggest that these isoforms may be important in odontoblast behaviour and the modulation of the tissue response to injury.

Edetic Acid↗
PubMed2003

Effect of hydrolyzed surface layer on the cytotoxicity and chemical resistance of a low fusing porcelain.

OBJECTIVE: The objectives of this study were to verify the formation of a hydrolyzed surface layer on Duceram LFC and to determine its effect on the cytotoxicity of the porcelain as measured by cellular activity and concentrations of leached ionic species. METHODS: Specimens were fabricated from dentin porcelain by a vibration blotting technique. Half of the specimens underwent accelerated aging by subjecting them to the standard test for hydrolytic resistance (ISO 6872). Fibroblast cell cultures were placed in direct contact with specimens. Cell viability was assessed using succinic dehydrogenase activity. Chemical resistance was determined by leaching specimens with a continuous flow of deionized water and measuring the concentrations of soluble ions in the leachates. Porcelain surface topography was examined using atomic force microscopy, and Fourier transform infrared spectroscopy was used to detect the composition of the surface layer. RESULTS: Hydrolyzation treatment created a smooth texture on the porcelain surfaces but did not result in a hydrolyzed surface layer with increased hydroxyl content. There was a decreased alkali ion content in the surface layer of hydrolyzed specimens. Mean cellular SDH activities for non-hydrolyzed and hydrolyzed porcelain were 75+/-7 and 80+/-5% of Teflon controls, respectively. Only sodium ions were present in significant concentrations in collected leachates. The sodium concentration decreased over the initial 4.5h of leaching. SIGNIFICANCE: The changes in surface layer texture and composition as the result of the aging treatment had little effect on the cytotoxicity and chemical resistance of Duceram LFC. The results suggest that Duceram LFC would pose no biocompatibility risk even after extended exposure to the oral environment. These data also provide baseline material properties to be used in future studies of the effects of porcelain additives.

3T3 Cells↗