scholarly journals Evaluation of the cytocompatibility of mixed bovine bone

2007 ◽  
Vol 18 (3) ◽  
pp. 179-184 ◽  
Author(s):  
Esther Rieko Takamori ◽  
Eduardo Aleixo Figueira ◽  
Rumio Taga ◽  
Mari Cleide Sogayar ◽  
José Mauro Granjeiro
Keyword(s):  
Cytotoxic Effect ◽  
Bone Matrix ◽  
Neutral Red ◽  
Water Soluble ◽  
Mechanical Resistance ◽  
Bovine Bone ◽  
3T3 Cells ◽  
Cell Based Therapy ◽  
Viable Cells

Treatment of bovine bone with peroxides and chaotropic agents aims to obtain an acellular bone matrix that is able to maintain the collagen-apatite complex and a higher mechanical resistance, a mixed biomaterial hereby named mixed bovine bone (MBB). The purpose of this study was to evaluate the cytocompatibility of MBB and cell-MBB interaction. Cell morphology, number of viable cells, ability to reduce methyltetrazolium and to incorporate neutral red upon exposure to different concentrations of the hydrosoluble extract of MBB were assessed in Balb-c 3T3 cells according to ISO 10993-5 standard. The interaction between cells and MBB surface was evaluated by scanning electron microscopy. The water-soluble MBB extracts were cytotoxic and led to cell death possibly due to its effect on mitochondrial function and membrane permeability. Cells plated directly onto the MBB did not survive, although after dialysis and material conditioning in DMEM + 10% FCS, the cells adhered and proliferated onto the material. It may be concluded that, in vitro, water-soluble MBB extracts were cytotoxic. Nevertheless, MBB cytotoxic effect was reverted by dialysis resulting in a material that is suitable for cell based-therapy in the bioengineering field.

scholarly journals Study of Two Bovine Bone Blocks (Sintered and Non-Sintered) Used for Bone Grafts: Physico-Chemical Characterization and In Vitro Bioactivity and Cellular Analysis

Materials ◽  
2019 ◽  
Vol 12 (3) ◽  
pp. 452 ◽  
Author(s):  
Sergio Gehrke ◽  
Patricia Mazón ◽  
Leticia Pérez-Díaz ◽  
José Calvo-Guirado ◽  
Pablo Velásquez ◽  
...  
Keyword(s):  
Physicochemical Properties ◽  
Structural Characteristics ◽  
Mercury Porosimetry ◽  
Negative Control ◽  
Infrared Spectrometry ◽  
Mechanical Resistance ◽  
Bovine Bone ◽  
In Vitro Bioactivity ◽  
Cellular Analysis

In this work, the physicochemical properties and in vitro bioactivity and cellular viability of two commercially available bovine bone blocks (allografts materials) with different fabrication processes (sintered and not) used for bone reconstruction were evaluated in order to study the effect of the microstructure in the in vitro behavior. Scanning electron microscopy, X-ray diffraction, Fourier transform infrared spectrometry, mechanical resistance of blocks, mercury porosimetry analysis, in vitro bioactivity, and cell viability and proliferation were performed to compare the characteristics of both allograft materials against a synthetic calcium phosphate block used as a negative control. The herein presented results revealed a very dense structure of the low-porosity bovine bone blocks, which conferred the materials’ high resistance. Moreover, relatively low gas, fluid intrusion, and cell adhesion were observed in both the tested materials. The structural characteristics and physicochemical properties of both ceramic blocks (sintered and not) were similar. Finally, the bioactivity, biodegradability, and also the viability and proliferation of the cells was directly related to the physicochemical properties of the scaffolds.

In Vitro and In Vivo Evaluation of a Calcium Phosphate and Demineralized Bone Matrix Composite Biomaterial

2010 ◽  
Vol 5 ◽  
pp. 1-12 ◽  
Author(s):  
A. Rosenberg ◽  
Aliassghar Tofighi ◽  
N. Camacho ◽  
J. Chang
Keyword(s):  
Calcium Phosphate ◽  
Demineralized Bone Matrix ◽  
Bone Matrix ◽  
Negative Control ◽  
Water Soluble ◽  
Dense Area ◽  
Viscosity Modifier ◽  
Demineralized Bone

A new class of osteoconductive and osteoinductive combination biomaterials composed of calcium phosphate cement (CPC), demineralized bone matrix (DBM) and a water-soluble viscosity modifier were prepared and characterized in-vitro and in-vivo. In previous studies, a range of combinations formulations were tested in order to compare their performance characteristic. In-vitro characterization results show that the mechanical strength is decreased when the amount of DBM increases. However, DBM does not affect the CPC’s ability to set hard and convert to nanocrystalline apatitic calcium phosphate, which shares the chemical structure of natural bone as seen in x-ray diffraction. It is known that the DBM alone is osteoinductive. In-vivo osteoinductivity testing of the formulations in an intramuscular, athymic rat model demonstrated that the combination material is also osteoinductive. Two formulations were chosen for in-vivo efficacy testing based on the results of in-vitro and in-vivo characterization. These formulations were studied using rabbit critical-sized femoral core defect model. The formulations were composed of DBM with particle sizes of 250 to 710 μm, carboxymethyl-cellulose (CMC) as the viscosity modifier and weight percent compositions of 50% DBM/ 45% CPC/ 5% CMC and 60% DBM/ 30% CPC/ 10% CMC. Bone integration and healing was graded at 6, 12, and 24 weeks. The two formulations were compared to the gold standard autograft at 12 weeks and to an empty defect as the negative control at 24 weeks. Based on micro-computed topography (μCT), both formulations allowed for continuity of bone throughout the defect region at all time points. No differences in dense area fraction were seen between two formulations at 6 weeks (p = 0.8661). There was no significant statistical difference between the two formulations and autograft at 12 weeks (p = 0.2467). At 24 weeks, both formulations had significantly higher dense area fractions than empty controls (p = 0.0001). Histologically, the biology of the treatment areas appeared to have returned to normal by 24 weeks with CPC appearing to be the principal osteogenic inducer. In conclusion, these combinations of CPC and DBM offers significant advantages (handling, mechanical properties and osteoinductivity) over current DBM products and can be an effective alternative to autograft in healing of bone defects.

scholarly journals In vitro induction of cartilage-specific macromolecules by a bone extract.

1983 ◽  
Vol 97 (6) ◽  
pp. 1950-1953 ◽  
Author(s):  
S M Seyedin ◽  
A Y Thompson ◽  
D M Rosen ◽  
K A Piez
Keyword(s):  
Morphological Changes ◽  
Bone Matrix ◽  
Type Ii Collagen ◽  
Enzyme Linked Immunosorbent Assay ◽  
Bovine Bone ◽  
In Vitro System ◽  
In Vitro Induction ◽  
Inhibition Technique

An in vitro system has been developed to study the onset of chondrogenesis. Embryonic rat muscle mesenchymal cells, when treated in suspension culture with an extract of bovine bone matrix, synthesized cartilage-specific proteoglycan and type II collagen. The synthesis of these two macromolecules was assayed by the enzyme-linked immunosorbent assay inhibition technique. Further evidence of chondrogenesis was demonstrated by morphological changes of treated cells when cultured in firm agarose and stained for metachromatic matrix. Even with crude bone matrix extracts, the assay was sensitive at the microgram level and significant differences in cartilage macromolecules compared with controls were observed in 2-3 d. In vivo the same extract induced first cartilage and then bone.

Cytotoxic Effect of Alcohol on Liver Cells and Fibroblasts in Vitro

1974 ◽  
Vol 19 (3) ◽  
pp. 125-127 ◽  
Author(s):  
F. Walker ◽  
W. Elmslie ◽  
R. A. Fraser ◽  
P. E. Snape ◽  
G. C. M. Watt
Keyword(s):  
Cell Survival ◽  
Liver Cell ◽  
Cytotoxic Effect ◽  
Liver Cells ◽  
Alcoholic Beverages ◽  
Pure Ethanol ◽  
Viable Cells

Cultures of liver cells (Chang) and of fibroblasts (3T6) were exposed to media containing a range of concentrations of ethanol. At the end of a standard period of time the number of viable cells was determined. As the concentration of ethanol increased over 0.25 per cent v/v liver cell survival decreased progressively. Fibroblast survival decreased progressively at ethanol concentrations over 0.50 per cent v/v. This indicates that ethanol is more toxic to liver cells than to fibroblasts. Certain alcoholic beverages, particularly brandy, were found to be more toxic than pure ethanol.

scholarly journals Bone Matrix Proteins: Isolation and Characterization of a Novel Cell-binding Keratan Sulfate Proteoglycan (Osteoadherin) from Bovine Bone

1998 ◽  
Vol 141 (3) ◽  
pp. 839-847 ◽  
Author(s):  
Mikael Wendel ◽  
Yngve Sommarin ◽  
Dick Heinegård
Keyword(s):  
Guanidine Hydrochloride ◽  
Bone Matrix ◽  
Keratan Sulfate ◽  
Apparent Size ◽  
Enzyme Linked Immunosorbent Assay ◽  
Bovine Bone ◽  
Metabolic Labeling ◽  
Cell Binding ◽  
Isolation And Characterization

A small cell-binding proteoglycan for which we propose the name osteoadherin was extracted from bovine bone with guanidine hydrochloride–containing EDTA. It was purified to homogeneity using a combination of ion-exchange chromatography, hydroxyapatite chromatography, and gel filtration. The Mr of the proteoglycan was 85,000 as determined by SDS-PAGE. The protein is rich in aspartic acid, glutamic acid, and leucine. Two internal octapeptides from the proteoglycan contained the sequences Glu-Ile-Asn-Leu-Ser-His-Asn-Lys and Arg-Asp-Leu-Tyr-Phe-Asn-Lys-Ile. These sequences are not previously described, and support the notion that osteoadherin belongs to the family of leucine-rich repeat proteins. A monospecific antiserum was raised in rabbits. An enzyme-linked immunosorbent assay was developed, and showed the osteoadherin content of bone extracts to be 0.4 mg/g of tissue wet weight, whereas none was found in extracts of various other bovine tissues. Metabolic labeling of primary bovine osteoblasts followed by immunoprecipitation showed the cells to synthesize and secrete the proteoglycan. Digesting the immunoprecipitated osteoadherin with N-glycosidase reduced its apparent size to 47 kD, thus showing the presence of several N-linked oligosaccharides. Digestion with keratanase indicated some of the oligosaccharides to be extended to keratan sulfate chains. In immunohistochemical studies of the bovine fetal rib growth plate, osteoadherin was exclusively identified in the primary bone spongiosa. Osteoadherin binds to hydroxyapatite. A potential function of this proteoglycan is to bind cells, since we showed it to be as efficient as fibronectin in promoting osteoblast attachment in vitro. The binding appears to be mediated by the integrin αvβ3, since this was the only integrin isolated by osteoadherin affinity chromatography of surface-iodinated osteoblast extracts.

Use of External Metabolising Systems in the Neutral Red Assay

1992 ◽  
Vol 20 (2) ◽  
pp. 262-265
Author(s):  
Eva Rasmussen
Keyword(s):  
Cell Lines ◽  
Cho Cells ◽  
Cytotoxic Effect ◽  
Neutral Red ◽  
3T3 Cells ◽  
Active Metabolites ◽  
Neutral Red Assay ◽  
Viability Assay ◽  
Toxic Metabolites ◽  
Mixed Function Oxygenase

BALB/C 3T3 mouse fibroblasts or CHO cells grown in 96-well microtitre plates were used for the neutral red viability assay. The cytotoxicity and ability of microsomes or an S9 mixed function oxygenase to activate cyclophosphamide were compared. A pronounced cytotoxic effect of both activation systems was found using CHO cells, but not using 3T3 cells. 3T3 cells were more sensitive than CHO cells to the active metabolites of cyclophosphamide. Cyclophosphamide was more efficiently converted to toxic metabolites by microsomes than by the S9 mix. A concentration of microsomes corresponding to 0.032mg protein per ml of medium was shown to be optimal for the activation of cyclophosphamide in both cell lines.

scholarly journals New Class of Anti-Inflammatory Therapeutics Based on Gold (III) Complexes in Intestinal Inflammation–Proof of Concept Based on In Vitro and In Vivo Studies

2021 ◽  
Vol 22 (6) ◽  
pp. 3121
Author(s):  
Julia B. Krajewska ◽  
Jakub Włodarczyk ◽  
Damian Jacenik ◽  
Radzisław Kordek ◽  
Przemysław Taciak ◽  
...  
Keyword(s):  
Mouse Model ◽  
Intestinal Inflammation ◽  
Therapeutic Potential ◽  
Gastrointestinal Diseases ◽  
Neutral Red ◽  
Water Soluble ◽  
In Vivo Studies ◽  
Anti Inflammatory

Inflammatory bowel diseases (IBD) are at the top of the worldwide rankings for gastrointestinal diseases as regards occurrence, yet efficient and side-effect-free treatments are currently unavailable. In the current study, we proposed a new concept for anti-inflammatory treatment based on gold (III) complexes. A new gold (III) complex TGS 121 was designed and screened in the in vitro studies using a mouse macrophage cell line, RAW264.7, and in vivo, in the dextran sulphate sodium (DSS)-induced mouse model of colitis. Physicochemical studies showed that TGS 121 was highly water-soluble; it was stable in water, blood, and lymph, and impervious to sunlight. In lipopolysaccharide (LPS)-stimulated RAW264.7 cells, the complex showed a potent anti-inflammatory profile, as evidenced in neutral red uptake and Griess tests. In the DSS-induced mouse model of colitis, the complex administered in two doses (1.68 μg/kg, intragastrically, and 16.8 μg/kg, intragastrically, once daily) produced a significant (* p < 0.05) anti-inflammatory effect, as shown by macroscopic score. The mechanism of action of TGS 121 was related to the enzymatic and non-enzymatic antioxidant system; moreover, TGS 121 induced changes in the tight junction complexes expression in the intestinal wall. This is the first study proving that gold (III) complexes may have therapeutic potential in the treatment of IBD.

Factors influencing synthesis and mineralization of bone matrix from fetal bovine bone cells grown in vitro

2009 ◽  
Vol 7 (7) ◽  
pp. 727-741 ◽  
Author(s):  
S. William Whitson ◽  
Marcia A. Whitson ◽  
Daniel E. Bowers ◽  
Michael C. Falk
Keyword(s):  
Bone Cells ◽  
Bone Matrix ◽  
Bovine Bone ◽  
Factors Influencing ◽  
Fetal Bovine

scholarly journals In vivo transdifferentiation, osteoconductive and osteoinductive properties of experimental water-soluble organo-biomaterials – A Pilot Study

2021 ◽  
Vol 10 (5) ◽  
pp. e45310515017
Author(s):  
Viviane Rozeira Crivellaro ◽  
Gilvan Spada ◽  
Cláudia Salete Judachesci ◽  
Paula Porto Spada ◽  
Luiza Rodrigues Saling ◽  
...  
Keyword(s):  
Bone Matrix ◽  
Negative Control ◽  
Water Soluble ◽  
Bovine Bone ◽  
Longitudinal Effects ◽  
Variance Homogeneity ◽  
Immunohistochemical Analyses ◽  
Control Sham

Inorganic bovine bone matrix (IBBM) is a biomaterial with proven osteoconductive functionalities. The objective of this study was to assess the in vivo bone regeneration functionalities of IBBM modified or not by an experimental MOE in sheep. MOE synthesis was performed by suspending nacre particles (0.05 g, diameters < 0.01 mm) in anhydrous acetic acid (pH 7, 5 mL, 25°C, 72 hours) using magnetic stirring. Polyethylene carriers (d= 5.0 mm, l= 10.0 mm, open ends) of negative control (sham) or experimental groups (IBBM or MOE-modified IBBM) were placed (n=3 conditions /animal; intramuscularly) adjacent to the lower spine of adult sheep (8 animals, » 45 Kg, 2 years old). Tissues were harvested (at 3 or 6 months) after implantation in preparation for histological (H), morphometrical (MM) and immunohistochemical analyses (IH; Wnt-3a, CD34, Vimentin and PREF-1). MM data were tested for normality and variance homogeneity using the Shapiro-Wilk and Levene tests, and Mann Whitney and Kruskal-Wallis, respectively. IM data were analyzed using two-way ANOVA and Tukey tests. Differences (p < 0.05) were observed between experimental groups (IBBM and IBBM+MOE at both 3 and 6 months) and controls (sham) for total area; Differences were not found for presence of remnant particles among experimental groups. The highest formation of bone was observed with IBBM+MOE (6-months). No differences (p > 0.05) were found on IM analysis (CD34, Vimentin, PREF-1, Wnt3a). Results indicated that experimental materials (IBBM+MOE) display promising functionalities. Additional studies are necessary to define biomaterials’ longitudinal effects and long-term biocompatibility properties.

scholarly journals Dynamics of reparative histogenesis of bone tissue in presence of some osteoplastic materials in vitro

2020 ◽  
Vol 4 (34) ◽  
pp. 46-50
Author(s):  
S. Yu. Ivanov ◽  
A. V. Volkov ◽  
D. A. De
Keyword(s):  
Bone Regeneration ◽  
Bone Tissue ◽  
Bone Matrix ◽  
Three Dimensional ◽  
Bone Defects ◽  
Control Group ◽  
Bovine Bone ◽  
Reparative Osteogenesis ◽  
The Impact

Currently, to solve the bone deficiency problem in the maxillofacial region, osteoplastic materials based on allogeneic and xenogenic collagen bone matrix are used, both in pure and in activated forms, by adding growth factors. It is impossible to determine the effectiveness and mechanisms of the osteoplastic materials effect on bone regeneration without a comprehensive study, including not only histological, but also morphometric studies of the structural components and cellular reactions in the impact area. Such studies provide reliable and objective information on the main processes taking place in bone regeneration.Purpose. To determine the spatial distribution of reparative osteogenesis in the presence of some osteoplastic materials in vitro.Materials and methods. Svetlogorsk breed pigs were used as a biomodel. Depending on the osteoplastic preparations used, the animals were divided into four groups of the two in each: 1st — a preparation based on a natural bovine bone graft was injected into bone defects. 2nd — a preparation based on collagenized porcine transplant was injected into bone defects. 3rd — a preparation consisting of 60 % hydroxyapatite (HA) and 40 % beta-tri-calcium phosphate; 4th — control group — the bone defect healed under a blood clot. Animals were removed from the experiment on the 45th day. We examined sections with a thickness of 20 μm using the method of light and fluorescence microscopy.Results. The results indicate different dynamics of the reparative osteogenesis in the presence of osteoplastic materials of different classes. In group 1, the filling of the defect with newly formed bone tissue is not uniform; in group 2, the filling of the defect with newly formed bone tissue is uniform; in group 3 the filling of the defect with non-formed bone tissue is uneven due to the pronounced hyperostosis; in the control group, the filling of the defect with newly formed bone tissue is not happening.Conclusion. Stimulation, the dynamics of reparative osteogenesis and the three-dimensional organization of bone regenerate depend on the osteoplastic material class, which requires further study of the dynamics and three-dimensional organization of bone regenerate to select the optimal bone-replacing agent.

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