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Biomedical subjects

W Geurtsen

Publications and source records attributed to W Geurtsen.

At least 37 records · Page 2Linked to original sources

Effects of mechanical force on primary human fibroblasts derived from the gingiva and the periodontal ligament.

Previous experiments have shown that mechanical stress may alter the interactions between cells and extracellular matrix (ECM). The purpose of our study was to investigate the effects of mechanical load on metabolism and ECM expression of primary human periodontal cells. The influence of gravitational force on proliferation, lactate dehydrogenase (LDH) release, and tenascin expression of gingival (HGF) and periodontal ligament fibroblasts (HPDL), as well as their adhesion to various extracellular matrix (ECM) components, was determined. Cells were centrifuged in microplates or flat tubes for 16 hrs at 217 g. Neither an enhanced release of LDH nor an alteration of cell proliferation could be detected after centrifugation. However, the attachment of loaded gingival and periodontal ligament fibroblasts to all tested ECM components significantly decreased in comparison with controls (Wilcoxon-Mann-Whitney test; HGF, p < 0.05; HPDL, p < 0.01). Tenascin expression of mechanically stressed fibroblasts significantly increased in comparison with controls (p < 0.01).

Cell Adhesion↗

Molar fracture resistance after adhesive restoration with ceramic inlays or resin-based composites.

PURPOSE: To determine the fracture resistance of teeth, following treatment with various types of adhesive restorations. MATERIALS AND METHODS: 5 0 caries-free, extracted human molars were randomly divided into five groups consisting of 10 molars each. MOD cavities were prepared in 40 molars with a width in the facio-lingual direction of 50% of the intercuspal distances. The cavities were filled with the following materials: Cerec or IPS Empress ceramic inlays, Arabesk or Charisma F resin-based composite (RBC) restorations. The control group consisted of 10 sound, non-restored molars. All 50 teeth were loaded occlusally until fracture using a tensile testing machine. The statistical analysis included ANOVA, Kolmogorov-Smimov-test, Scheffé test, and boxplots. RESULTS: There was no significant difference (P > 0.05) between the mean values of the sound teeth (2,102 N) and the teeth with the Cerec ceramic inlays (2,139 N). However, both groups demonstrated a significant difference (P < 0.05) when compared with the teeth with IPS Empress ceramic inlays (1,459 N) and Arabesk RBC restorations (1459 N). No significant differences were found between the last two groups. Molars restored with Charisma F composite restorations (1,562 N) revealed no significant difference when compared with all other groups including controls (P > 0.05). A stabilization of molars is possible by means of an adhesive restoration in the form of an "internal splinting" regardless of the restorative material used.

Acrylates↗

Effects of BisGMA and TEGDMA on proliferation, migration, and tenascin expression of human fibroblasts and keratinocytes.

Previous studies have documented a marked cytotoxic potency of BisGMA and TEGDMA. The purpose of this investigation was to determine if these substances also affect proliferation, migration, and tenascin expression of primary human gingival fibroblasts (HGF) and immortalized human keratinocytes (HaCaT). These parameters play an important role in healing wounds. HGF and HaCaT cultures were incubated with TEGDMA and BisGMA. Cell proliferation (BrdU-assay) and migration (Boyden method) were determined 24 h after incubation. Tenascin expression was investigated four and seven days after treatment. Results were statistically evaluated by ANOVA using the Wilcoxon-Mann-Whitney test (p < 0.05). Proliferation of both cell types was significantly inhibited at concentrations > or = 0.25 mM (TEGDMA) or > or = 0.01 mM (BisGMA). Migration of HaCaT was significantly increased after incubation with BisGMA for 24 h. TEGDMA did not alter migration of HGF and HaCaT. In addition, TEGDMA had no effect on tenascin expression of both cell cultures. After 4 days of incubation, BisGMA (at a concentration of 0.01 mM) significantly reduced tenascin production of HaCaT cultures related to cell number. However, 7 days after treatment, BisGMA significantly increased tenascin expression of HGF and HaCaT cultures. Altogether, our results indicate that BisGMA can affect migration of keratinocytes and alters the expression of the extracellular matrix component tenascin. Thus, BisGMA may significantly influence the healing of injured oral tissues.

Antimetabolites↗

Compatibility of resorbable and nonresorbable guided tissue regeneration membranes in cultures of primary human periodontal ligament fibroblasts and human osteoblast-like cells.

The purpose of this study was (a) to evaluate the cytocompatibility of three resorbable and nonresorbable membranes in fibroblast and osteoblast-like cell cultures and (b) to observe the growth of those cells on the various barriers by scanning electron microscopy (SEM). Primary human periodontal ligament fibroblasts (HPLF) and human osteoblast-like cells (SAOS-2) were incubated with nonresorbable polytetrafluoroethylene (ePTFE) barriers and resorbable polylactic acid as well as collagen membranes. Cytotoxic effects were determined by XTT (mitochondrial metabolic activity) and sulforhodamine B assays (cellular protein content). In addition, HPLF and SAOS-2 grown for 21 days on the investigated barriers were evaluated by SEM. Data were analyzed statistically by ANOVA using the Wilcoxon-Mann-Whitney test (P < 0.05). No changes were established in the periodontal ligament fibroblasts and human osteoblast-like cells after incubation with the collagen membrane. Cytotoxic effects, however, were induced by the polylactic acid barrier which slightly inhibited cell metabolism of the periodontal fibroblasts (XTT: 90.1% +/- 3.6 of control value). Moderate cytotoxic reactions were caused by the nonresorbable ePTFE membrane in HPLF-cultures (XTT: 82.7% +/- 3.5) and osteoblast-like cell monolayers (XTT: 80.0% +/- 0.6%). Mitochondrial activity in both cell cultures was significantly reduced by ePTFE barriers in comparison to non-incubated control cells (P = 0.028). SEM analysis of cell behavior on barriers demonstrated the differences between these materials: collagen barriers were densely populated with HPLF and SAOS-2, whereas only few or no cells were seen to adhere to the ePTFE and polylactic acid membranes. Our findings indicate that the collagen barrier investigated is very cytocompatible and may be integrated into connective tissue well. On the contrary, the ePTFE and polylactic acid membranes induced slight to moderate cytotoxic reactions which may reduce cellular adhesion. Thus, gap formation between the barrier surface and the connective tissue may be promoted which may facilitate epithelial downgrowth and microbial accumulation. Consequently, these effects may reduce the potential gain in periodontal attachment.

Absorbable Implants↗

Infection of primary human gingival fibroblasts by Porphyromonas gingivalis and Prevotella intermedia.

Adhesion and penetration of clinical isolates of Porphyromonas gingivalis and Prevotella intermedia in human gingival fibroblast monolayers were studied by transmission electron microscopy (TEM). Fibroblasts were cultured from biopsies of human healthy gingiva. Porphyromonas gingivalis and Prevotella intermedia were isolated from patients with periodontitis. Fibroblasts were incubated with microorganisms in an antibiotic-free medium for 24 h. Then cultures were washed to remove nonadherent bacteria. Consecutively, infected cultures were grown for another 24 h. Thereafter, the treated monolayers were prepared for TEM investigations. Internalized Porphyromonas gingivalis and Prevotella intermedia were visible after 24 h of incubation. Prevotella intermedia showed only division in cytoplasm of fibroblasts after 24 h and 48 h incubations. Infected fibroblasts revealed various morphological alterations such as extensive vacuolization and breakdown of mitochondria. These findings demonstrate that Porphyromonas gingivalis and Prevotella intermedia may invade human gingival fibroblasts and thus may damage these cells directly or due to the release of microbial cytotoxic components.

Adult↗

Comparison of the number and diameter of dentinal tubules in human and bovine dentine by scanning electron microscopic investigation.

Detailed information on dentine structure is essential for interpreting data from investigations on dentine-adhesive materials. The purpose here was to compare the number and diameter of dentinal tubules at similarly prepared surfaces of bovine permanent central incisors and human deciduous and third molars. In bovine teeth, crowns and roots were used; in human samples only the crowns were investigated. Tubule density in the middle layer was higher in bovine root (BR) dentine (number of tubules per mm(2)+/-SD: 23, 760+/-2453) than in human deciduous (HD) (18,243+/-3845), human permanent (HP) (18,781+/-5855), and bovine coronal (BC) (17, 310+/-2140) dentine. The corresponding values for the deep layer were 23,738+/-4457 (BR), 24,162+/-5338 (HD), 21,343+/-7290 (HP), and 20,980+/-4198 (BC). No significant differences were found for the number of dentinal tubules in bovine coronal dentine compared to the dentine of human deciduous and permanent molars. The mean diameter of bovine dentinal tubules was slightly, but not significantly, higher than in human dentine (middle layer/deep layer+/-SD): BC, 2. 85 microm+/-0.18/3.50 microm+/-0.33; BR, 3.10 microm+/-0.33/3.23 microm+/-0.30; HD, 2.55 microm+/-0.16/2.82 microm+/-0.28; HP, 2.65 microm+/-0.19/2.90 microm+/-0.22. These findings demonstrate that corresponding coronal dentine layers of human deciduous and permanent molars, and of bovine central incisors, are not significantly different in their number of tubules per mm(2) and their tubule diameter, whereas tubule density in bovine root dentine is significantly higher. These results suggest that provided standardized preparations are used, bovine incisor crown dentine is a suitable substitute for human molar dentine in adhesion studies.

Adolescent↗

Biocompatibility of resin-modified filling materials.

Increasing numbers of resin-based dental restorations have been placed over the past decade. During this same period, the public interest in the local and especially systemic adverse effects caused by dental materials has increased significantly. It has been found that each resin-based material releases several components into the oral environment. In particular, the comonomer, triethyleneglycol di-methacrylate (TEGDMA), and the 'hydrophilic' monomer, 2-hydroxy-ethyl-methacrylate (HEMA), are leached out from various composite resins and 'adhesive' materials (e.g., resin-modified glass-ionomer cements [GICs] and dentin adhesives) in considerable amounts during the first 24 hours after polymerization. Numerous unbound resin components may leach into saliva during the initial phase after polymerization, and later, due to degradation or erosion of the resinous restoration. Those substances may be systemically distributed and could potentially cause adverse systemic effects in patients. In addition, absorption of organic substances from unpolymerized material, through unprotected skin, due to manual contact may pose a special risk for dental personnel. This is borne out by the increasing numbers of dental nurses, technicians, and dentists who present with allergic reactions to one or more resin components, like HEMA, glutaraldehyde, ethyleneglycol di-methacrylate (EGDMA), and dibenzoyl peroxide (DPO). However, it must be emphasized that, except for conventional composite resins, data reported on the release of substances from resin-based materials are scarce. There is very little reliable information with respect to the biological interactions between resin components and various tissues. Those interactions may be either protective, like absorption to dentin, or detrimental, e.g., inflammatory reactions of soft tissues. Microbial effects have also been observed which may contribute indirectly to caries and irritation of the pulp. Therefore, it is critical, both for our patients and for the profession, that the biological effects of resin-based filling materials be clarified in the near future.

Animals↗

Placement of the preparation line and periodontal health--a prospective 2-year clinical study.

The aim of this prospective clinical 2-year study was to determine whether the placement of the proximal margins of crowns within the zone of the biologic width results in periodontal alterations. In 41 patients, 116 prepared teeth as well as 82 unrestored, healthy contralateral teeth were examined. The following indices were determined before as well as 3, 6, 12, and 24 months after preparation: hygiene index, papillary bleeding index (PBI), and probing depth. After preparation, the distance between the restoration margins and the alveolar crest was measured with a modified periodontal probe on both proximal aspects of each tooth. These values were classified into 3 groups: I = < or = 1 mm between crown margin and alveolar crest, II = 1 to 2 mm, and III = > 2 mm. In addition, radiographs were taken directly after cementation of each restoration and after 12 and 24 months. The hygiene index did not significantly vary during the investigation, but PBI increased in all groups. The highest PBI increase was found in group I; in this group, the most significant increase was found between 3 and 6 months after preparation. The mean probing depth only increased in group I during the first 3 months after preparation on the mesial aspects. Only minor changes were found on the distal aspects of the teeth in group I and on all aspects in groups II and III. No alterations of the bone levels were diagnosed on the radiographs. These results indicate that the location of the restoration margins within the zone of the biologic width may impair the periodontal health of restored teeth.

Adult↗

Rapid general dental erosion by gas-chlorinated swimming pool water. Review of the literature and case report.

Several reports indicate an increased prevalence of dental erosion among intensive swimmers due to low pH gas-chlorinated pool water. Contrary to other extrinsic factors which induce erosion located on the facial aspect, low pH pool water results in general dental erosion. Additionally, a case report is presented which describes the very rapid occurrence of excessive general dental erosion of a competitive swimmer due to gas-chlorinated pool water within 27 days. The observation of several authors as well as this case underscore the significance of a regular pH monitoring of chlorinated swimming pool water. The high incidence indicates that dental erosion due to frequent swimming is of considerable diagnostic and therapeutic significance. Furthermore, it is recommended to fluoridate the teeth of intensive swimmers regularly to prevent dental erosion.

Adult↗

Aqueous extracts from dentin adhesives contain cytotoxic chemicals.

It was the aim of our study to investigate the composition and cytotoxicity of aqueous elutes from five dentin adhesives currently used in clinical practice: Solobond Plustrade mark, Solisttrade mark, Scotchbond Multipurposetrade mark, Syntac SCtrade mark, and Prime & Bondtrade mark 2.1. Water extracts were analyzed by gas chromatography/mass spectrometry (GC/MS) and relative quantities of identified compounds were compared by means of an internal caffeine standard [%CF]. The in vitro cytotoxic effects of substances released into DMEM were determined using immortalized 3T3-fibroblast cultures. In addition, the cytotoxicity of ethylene glycol (EG), which was identified in the extracts of Syntac SC, was evaluated. All dentin adhesives tested released various chemical components, like comonomers (mainly ethylene glycol compounds), HEMA, and initiating substances (e.g., camphorquinone). Elutes of Solobond Plus, which contained very high amounts of TEGDMA, were extremely cytotoxic. Two bonding agents (Scotchbond Multi-purpose, Syntac SC), which released significant quantities of HEMA, induced severe cytotoxic effects. In contrast, extracts from Solist and Prime & Bond 2.1 had very small effects on cell proliferation; these elutes contained small amounts of released chemical compounds. EG, a product of HEMA hydrolysis, in concentrations ranging from 0.025-25 mM was not cytotoxic. In summary, these results provide evidence that all dentin adhesives tested in the present study release in aqueous media chemical compounds some of which (for example, TEGDMA and HEMA) are cytotoxic.

3T3 Cells↗

Variability of cytotoxicity and leaching of substances from four light-curing pit and fissure sealants.

It was the aim of our study to investigate the composition and cytotoxicity of aqueous extracts of four light-curing pit and fissure sealants. Water extracts were analyzed by gas chromatography/mass spectrometry (GC/MS), and relative quantities of identified compounds were compared by means of an internal caffeine standard [%CF]. Cytotoxic effects due to medium extracts were determined by means of permanent 3T3 fibroblasts. All light-curing pit and fissure sealants segregated different ingredients into water, such as co-monomers (mainly ethylene glycol compounds) and initiating substances (e.g., camphorquinone). Bisphenol-A, however, which is easily detected by GC/MS, was not found in any of the analyzed eluates. The extracts of three sealants inhibited monolayer growth only moderately whereas the eluate of one product inhibited cell proliferation significantly. In the extracts of this sealant high quantities [%CF] of the co-monomer TEGDMA were detected. Our results indicate that light-curing pit and fissure sealants release substances into aqueous media that may induce cytotoxic effects. However, no concerns about potential estrogenic effects of Bisphenol-A are supported by our results.

3T3 Cells↗

Cytocompatibility of periodontal dressing materials in fibroblast and primary human osteoblast-like cultures.

Purpose of this investigation was to determine the cytocompatibility of various periodontal dressing materials by means of human primary gingival fibroblasts (HGF), human osteoblast-like cells (HObl) derived from the alveolar bone, and permanent 3T3 mouse fibroblasts (3T3). Cell culture medium extracts (time periods of extraction: day 1 and between day 2 and day 8 after setting) as well as solid specimens of the following materials were investigated: Coe-pak, Voco pac, Peripac, and Barricaid. Responses of cultures exposed for 24 h and 48 h to these materials were monitored by the fluorescent dyes H33342 and sulforhodamin 101 as well as by light microscopy. It was found that most extracts of Voco pac, Peripac, and Barricaid did not inhibit growth of HGF. Coe-Pak, however, clearly reduced the proliferation of HGF compared to untreated controls. Peripac decreased growth of HObl whereas Coe-Pak, Voco pac, and Barricaid caused no cytotoxic alterations in any of the test assays. Contrary to HGF and HObl, 3T3 cells were much more irritated by the test materials. But the light-curing resinous material Barricaid reduced proliferation of 3T3-fibroblasts only slightly. Our data indicate that Barricaid is exceedingly cytocompatible, whereas all other materials revealed moderate or severe cytotoxic effects according to the cell type.

3T3 Cells↗

Genotoxicity and cytotoxicity of the epoxy resin-based root canal sealer AH plus.

Previous studies with four rapid in vitro and in vivo test systems have shown that the epoxy resin-based root canal sealer AH26 may be genotoxic and cytotoxic (9). The purpose of this study was to determine the cytotoxic and genotoxic effects of the new resinous root canal sealer AH Plus by means of the growth inhibition test with primary human periodontal ligament fibroblasts and permanent 3T3 monolayers, the procaryotic umu test, the eucaryotic DNA synthesis inhibition test, and the in vivo alkaline filter elution test. In addition, Ames tests were performed with extracts from AH Plus. AH Plus caused only slight or no cellular injuries. Furthermore, no genotoxicity and mutagenicity were revealed by AH Plus. These data should be taken into consideration when deciding about a root canal sealer.

3T3 Cells↗

Effect of storage media on the fluoride release and surface microhardness of four polyacid-modified composite resins ("compomers").

OBJECTIVES: The aims of this investigation were to measure the surface microhardness (Vickers) as well as the release of fluoride from four polyacid-modified composite resins (PMC) ("compomers") (Compoglass F, F 2000, Dyract AP, experimental compomer) after storage in various artificial saliva (buffers) including one esterase-buffer. METHODS: Samples were stored for 6 days in de-ionized water, acidic buffer I (pH 4.2), neutral buffer II (pH 7.0), or neutral buffer III (pH 7.0) containing porcine esterase. The specimens were transferred into fresh media every 48 h. Fluoride release was measured every 48 h. Vickers hardness of each five samples of every group was determined before storing the samples in media (baseline) as well as after storage for 24, 48, and 144 h in the various solutions. Dry-stored specimens served as control. RESULTS: The surface microhardness of all PMCs significantly decreased after storage in the various media. No significant differences, however, were found between samples of the same material stored in the various media for 6 days. In general, the highest fluoride quantity was released into the acidic buffer I except for Dyract AP, which segregated similar quantities of fluoride into buffer I and into de-ionized water. More fluoride was released into de-ionized water than into neutral buffers. Further, esterase treatment increased fluoride release from three PMCs. SIGNIFICANCE: Our results suggest that the action of salivary esterases may weaken the surface of polyacid-modified composite resin restorations. As a clinical consequence, wear may be enhanced and load resistance may be reduced. In addition, fluoride release from PMCs may be increased by hydrolytic enzymes in saliva and under acidic conditions.

Analysis of Variance↗

Matrix expression and proliferation of primary gingival fibroblasts in a three-dimensional cell culture model.

The growth of cultured primary human gingival fibroblasts and the three-dimensional arrangement of the extracellular matrix in a polyester carrier system was investigated using various histological techniques. The results were compared with monolayer cultures. Collagen types I, III, V, and VI were investigated by conventional and fluorescence microscopy, scanning and transmission electron microscopy, and confocal laser scanning microscopy. Human gingival fibroblasts were obtained from tissue biopsies of five donors and were cultivated up to 5 weeks under three-dimensional culture conditions. The cells displayed an elongated, spindle-like or stellate morphology resembling the in vivo situation. Collagen type I revealed thick fiber bundles, and collagens type III and V were distributed as fine fibrils or small bundles throughout the culture system. Frequently, the fibers were oriented parallel to the long axis of the cells. Type VI collagen formed thin fibers and revealed a reticular pattern. In histological sections the cultured cells exhibited a morphology clearly different from that of cells cultured in monolayers. Their shape and spatial distribution resembled that of cells in tissue biopsies more closely. The culture system presented here promotes a dynamic model for performing studies for instance on the interactions of cultured cells with extracellular matrix molecules, on the pathogenesis of inflammatory processes or on the interactions with biomaterials, thus providing qualitative and quantitative information.

Adolescent↗

Bovine dentin as a substitute for human dentin in shear bond strength measurements.

PURPOSE: To evaluate the suitability of bovine coronal and root dentin as substitutes for human primary and permanent dentin in shear bond strength tests for dentin adhesives. MATERIALS AND METHODS: 30 bovine permanent central incisors, 30 human primary and 30 human third molars were cut mesiodistally. The pulpal and buccal dentin surfaces of the buccal cuts were milled with a bur to a thickness of 1.0 mm. A dentin adhesive and a hybrid composite were applied exactly according to the instructions given by the manufacturer on each buccal and pulpal surface, except in primary teeth where only the buccal surface was used. Shear bond strength was determined after 24 hours of storage in an aqueous solution. The results were statistically analyzed using the Wilcoxon- or the Mann-Whitney-U-test. RESULTS: There were no differences in shear bond strengths between human permanent dentin and bovine coronal dentin (13.3 +/- 6.1 vs. 15.2 +/- 7.6), and between each pulpal and buccal aspect. Significant differences were found between bovine root dentin and human primary dentin (17.4 +/- 8.3 vs. 7.7 +/- 5.0, P < 0.001). Significant differences were also found between bovine root dentin and human permanent dentin (17.4 +/- 8.3 vs. 13.3 +/- 6.1, P < 0.05), and bovine root and coronal dentin (17.4 +/- 8.3 vs. 15.2 +/- 7.6, P < 0.05). Furthermore, significant differences were recorded between human primary and human permanent dentin (7.7 +/- 5.0 vs. 13.3 +/- 6.1, P < 0.001) and bovine coronal dentin (7.7 +/- 5.0 vs. 15.2 +/- 7.6, P < 0.001).

Animals↗

Bonded restorations for the prevention and treatment of the cracked-tooth syndrome.

Several reports revealed that the cracked-tooth syndrome is a common problem in dental practice, which often results in extraction of the affected incompletely fractured teeth. Predominantly restored teeth suffer from these incomplete fractures. Therefore, it is of outstanding importance to stabilize teeth weakened due to cavity preparation. Besides full cuspal coverage by partial or full crowns, bonded restorations have been proposed for internal splinting of restored teeth. Although contradictory data have been published, there is evidence that bonded amalgam or resin-based composite restorations (RBC) do not increase fracture resistance of teeth with wide occlusal-proximal cavities to values similar to sound, unrestored controls. Indirectly fabricated RBC inlays and various ceramic inlays, however, increased fracture strength to levels as high as those of sound caries-free teeth. Therefore, it is recommended that weakened teeth with wide cavities be strengthened by full cuspal coverage with cast or ceramic restorations, by bonded ceramic inlays, or by indirectly-fabricated bonded RBC composite inlays.

Composite Resins↗

Cytotoxicity of 35 dental resin composite monomers/additives in permanent 3T3 and three human primary fibroblast cultures.

It was the purpose of this investigation to determine the cytotoxic effects (ED50 concentrations) of 35 monomers or additives identified in commercial dental resin composites. Monolayers of permanent 3T3 cells and three primary human fibroblast types derived from oral tissues (gingiva, pulp, and periodontal ligament) were used as test systems. All substances were tested in concentrations ranging from 0.01 to 5.0 mM. In general, ED50 values varied from 0.06 to > 5 mM. Within the groups of co(monomers), initiators, and cointiators, severe (e.g., Bis-GMA, UDMA, DMBZ, and DMDTA) or moderate (HEMA, BEMA, CQ, DMPT, and DMAPE) cytotoxic effects could be evaluated. Within the group of reaction/decomposition products, only moderate or slight effects were found (ED50: 0.7 to > 5 mM). The inhibitor BHT, the contaminant TPSb, and the photostabilizer HMBP, however, were highly cytotoxic in all cell cultures. In addition, the ED50 values of DBPO and HMBP significantly varied (0.43-3.8 mM, respectively, and 0.44-3.07 mM) with the applied cell culture. Our comprehensive screening shows that for several of the highly cytotoxic composite components, less cytotoxic alternatives are available. Furthermore, there was no cell type identified which was consistently less or more sensitive to the toxic effects of the tested compounds than the others. Primary human periodontal ligament and pulp fibroblasts, however, were found to be more sensitive than 3T3 and gingival fibroblasts to alterations from most tested substances.

3T3 Cells↗