Background: Regarding retained cement acts as an idiopathic cause for early implant loss, cement ... more Background: Regarding retained cement acts as an idiopathic cause for early implant loss, cement selection with the least toxicity in the peri-implant hard and soft tissues is important. The present study aimed to evaluate the cytotoxicity of three different types of temporary cements and titanium cemented with each cement after direct exposure to human gingival fibroblasts (HGF) and MG-63 osteoblast-like cells.Methods: In this in vitro study, zinc oxide-eugenol (ZOE), eugenol-free zinc oxide (ZONE), and resin (R) cements prepared in cylindrical forms with a similar dimension were used. Each cement was placed on titanium disks to prepare cemented titanium samples. MTT assay was applied at 24- and 72-hour, and 7-day intervals to evaluate cytotoxicity.Results: All the cements decreased cell viability in both cell lines significantly. None of the cements exhibited cellular viability percentages higher than the minimum percentage (70%) required for biocompatibility. The cemented titaniu...
Abstract Context Bacillus anthracis secretes a tripartite toxin comprising protective antigen (PA... more Abstract Context Bacillus anthracis secretes a tripartite toxin comprising protective antigen (PA), edema factor (EF), and lethal factor (LF). The human anthrax vaccine is mainly composed of the anthrax protective antigen (PA). Considerable efforts are being directed towards improving the efficacy of vaccines because the use of commercial anthrax vaccines (human/veterinary) is associated with several limitations. Objective In this study, a triple chimeric antigen referred to as ELP (gene accession no: MT590758) comprising highly immunogenic domains of PA, LF, and EF was designed, constructed, and assessed for the immunization capacity against anthrax in a guinea pig model. Materials and methods Immunization was carried out considering antigen titration and immunization protocol. The immunoprotective efficacy of the ELP was evaluated in guinea pigs and compared with the potency of veterinary anthrax vaccine using a challenge test with B. anthracis 17JB strain spores. Results The results demonstrated that the ELP antigen induced strong humoral responses. The T-cell response of the ELP was found to be similar to PA, and showed that the ELP could protect 100%, 100%, 100%, 80% and 60% of the animals from 50, 70, 90, 100 and 120 times the minimum lethal dose (MLD, equal 5 × 105 spore/ml), respectively, which killed control animals within 48 h. Discussion and conclusions It is concluded that the ELP antigen has the necessary requirement for proper immunization against anthrax and it can be used to develop an effective recombinant vaccine candidate against anthrax.
Abstract Collagen as a biomaterial is commonly obtained from terrestrial animals. However, nowada... more Abstract Collagen as a biomaterial is commonly obtained from terrestrial animals. However, nowadays, the use of collagen with terrestrial animals' sources due to possible transmission of infectious diseases and religious beliefs is restricted. This study was conducted to evaluate the physicochemical characterization, morphology, and biocompatibility of extracted collagen from silver carp fish skin by-product. Type I collagen was extracted from the silver carp fish skin by-product using acetic acid and pepsin enzyme. The results showed that the extraction yield of ASC and PSC was 42.85% and 58.75% (on a dry weight basis), respectively. The presence of two different α-chains confirmed the type I collagen structure for both collagens. Moreover, FTIR spectra investigation demonstrated the triple helical in ASC and PSC collagens. The PSC showed higher imino acids content than ASC. Hence, the fractional viscosity and DSC curves revealed higher Td (30 °C), Ts (81 °C), and Tm (209 °C) for PSC than ASC (28.73 °C, 77 °C, and 187 °C, respectively). Finally, observations of the microscopic and the cell viability evaluation confirmed biocompatibility and suitable structure to cell growth. Accordingly, the obtained collagen from silver carp fish skin can be a proper alternative for terrestrial animals’ collagen and a safe biomaterial for biomedical use.
Abstract In the present study, Poly l -lactic acid (PLLA) resin compatible with digital light pro... more Abstract In the present study, Poly l -lactic acid (PLLA) resin compatible with digital light processing (DLP) 3D printing method was synthesized to produce hard tissue scaffolds. PLLA has been chosen as a decent material to mimic biological structures due to its relatively high strength as well as proper biocompatibility and biodegradation rate. After synthesis and functionalization of PLLA, using a facile method, porous models with 600-micron pore size and 70 % nominal porosity were designed and fabricated via DLP technique in order to investigate the effects of the two process parameters, light exposure time and dye concentration, on compressive strength and morphological features of the printed samples. The experimental results were then reconciled with plotted working curves for each dye concentration to validate the defined exposure time levels. It was concluded that the synthesized polymer and the used method of 3D printing are suitable for fabricating scaffolds with intricate structures. Moreover, by conducting the compression test, a maximum 2.2 MPa strength was achieved for the sample with minimum dye concentration and maximum exposure time. From the biological point of view, no cytotoxic effect was seen after a 3-day in vitro cell viability testing. Altogether, it was shown that optimal adjustment of the process parameters is essential to achieve appropriate dimensional and mechanical properties, which were acknowledged by plotted working curves.
1St Tabriz International Life Science Conference and 12th Iran Biophysical Chemistry Conference, May 22, 2013
Introduction : The PCR is an indispensable method in biotechnology. One of the limitations of PCR... more Introduction : The PCR is an indispensable method in biotechnology. One of the limitations of PCR is the size of the amplified DNA product. The basis for this constraint is the error rate and low proccessivity and elongation rate of different thermostable DNA polymerases. However, if an efficient thermostable PCR enzyme with higher accuracy, elongation velocity and proccessivity such as KOD DNA polymerase is used an increased yield of a high fidelity long_range PCR products can be produced. The aim of this study was to optimize of the production and purification of a functional KOD DNA polymerase. Method : We have purchased a pET17b plasmid that had KOD DNA polymerase gene in multiple cloning site and transform it in the expressing strain BL21(DE3) containing the pLysE or pLysS plasmid. Production of the 90-kDa protein was induced by different concentrations of IPTG. Purification were performed by heating, ammonium sulfate precipitation and chromatography with DEAE-sepharose. Purified protein was identified by SDS-PAGE and AgNO3 staining. The activity of the obtained enzyme was measured by comparing the intensities of the produced DNA bands in PCR reactions. Results: The correct framing of the gene and its orientation were analyzed by digestion and colony PCR. The plasmid was stable in BL21(DE3) containing the pLysS plasmid. Addition of glycerol, Triton X-100, Tween 20, dextrose and protease inhibitor cocktail increased enzyme stability. Conclusions: We have found that pET17b expression vector is toxic or unstable in the expressing strain BL21(DE3), even in the absence of induction. However BL21(DE3) containing the pLysS plasmid suppresses expression prior to induction and was able to produce greater amount of enzyme after induction by IPTG.
Artificial cells, nanomedicine, and biotechnology, Jan 26, 2018
Zein nanoparticles as a carrier system for BMP6-derived peptide were prepared by liquid-liquid ph... more Zein nanoparticles as a carrier system for BMP6-derived peptide were prepared by liquid-liquid phase separation procedure and characterized with SEM, DLS, FTIR and thermogravimetric methods. After peptide encapsulation, nanoparticle size increased from 236.3 ± 92.2 nm to 379.4 ± 116.8 nm. The encapsulation efficiency of peptide was 72.6% and the release of peptide from Zein nanoparticles was partly sustained in trypsin containing phosphate buffered saline (pH 7.4) for up to 14 days. Peptide-loaded nanoparticles showed similar cell viability compared with blank ones. ALP activity of C2C12 cells treated with peptide-loaded nanoparticles (500 µg/mL) was evaluated 7, 14, 21 and 28 days after culture. In peptide-loaded nanoparticles, ALP activity was significantly higher (p < .05) compared with other groups at day 14. Alizarin Red S staining showed, C2C12 cells behind peptide-loaded nanoparticles had significantly (p < .05) higher calcium deposition at day 21. The results of RT-qPC...
The design of a new truncated and engineered alpha1-antitrypsin based on theoretical studies: an ... more The design of a new truncated and engineered alpha1-antitrypsin based on theoretical studies: an antiprotease therapeutics for pulmonary diseases
Cholesteryl ester transfer protein (CETP) catalysis the exchange of triglycerides (TG) and choles... more Cholesteryl ester transfer protein (CETP) catalysis the exchange of triglycerides (TG) and cholesterol esters among plasma lipoproteins. It has been shown that variations at CETP locus is important in the levels and activity of CETP and high density lipoprotein (HDL) plasma concentration. In this research, we assessed the relationship between TaqIB CETP polymorphism and high density lipoprotein-cholestrol (HDL-C) concentration in a study sample of 128 Iranian residents. 1Based on our investigations, it was shown that presence of B1B1 genotype related to TaqIB polymorphism was negatively correlated with HDL-C and HDL2-C, but positively correlated with LDL-C. On the other hand, no correlation with HDL3-C was observed. Analysis of HDL-C subfraction in CHD and control subjects revealed that variation of the CETP TaqI B locus was significantly associated with concentration of HDL2-C subclass. Homozygous and heterozygous carried the B2 alleles, compared with B1 homozygote had significantl...
Human epidermal growth factor receptor 2 (HER2)-positive breast cancer represents approximately 1... more Human epidermal growth factor receptor 2 (HER2)-positive breast cancer represents approximately 15-30% of all invasive breast cancers. Despite the recent advances in therapeutic practices of HER2 subtype, drug resistance and tumor recurrence still have remained as major problems. Drug discovery is a long and difficult process, so the aim of this study is to find potential new application for existing therapeutic agents. Gene expression data for breast invasive carcinoma were retrieved from The Cancer Genome Atlas (TCGA) database. The normal and tumor samples were analyzed using Linear Models for Microarray Data (LIMMA) R package in order to find the differentially expressed genes (DEGs). These genes were used as entry for the library of integrated network-based cellular signatures (LINCS) L1000CDS2 software and suggested 24 repurposed drugs. According to the obtained results, some of these drugs including vorinostat, mocetinostat, alvocidib, CGP-60474, BMS-387032, AT-7519, and curcumin have significant functional similarity and structural correlation with FDA-approved breast cancer drugs. Based on the drug-target network, which consisted of the repurposed drugs and their target genes, the aforementioned drugs had the highest degrees. Moreover, the experimental approach verified curcumin as an effective therapeutic agent for HER2 positive breast cancer. Hence, our work suggested that some repurposed drugs based on gene expression data can be noticed as potential drugs for the treatment of HER2-positive breast cancer.
Due to the unique features of xenografts including large supply from donors, minimal risk of huma... more Due to the unique features of xenografts including large supply from donors, minimal risk of human disease transmission, and the lower cost of preparation and production compared to autografts and allografts, they are considered as attractive alternatives to traditional bone grafts. The animal source accessibility and production process have a direct correlation with the cost and quality of the final product. To evaluate whether the animal source of the bone has any effect on the physicochemical and histological properties of the final xenograft, three deproteinized bone grafts were prepared from sources that are easily available in Iran, including the bovine (DBB), camel (DCB), and ostrich (DOB).
Background: Regarding retained cement acts as an idiopathic cause for early implant loss, cement ... more Background: Regarding retained cement acts as an idiopathic cause for early implant loss, cement selection with the least toxicity in the peri-implant hard and soft tissues is important. The present study aimed to evaluate the cytotoxicity of three different types of temporary cements and titanium cemented with each cement after direct exposure to human gingival fibroblasts (HGF) and MG-63 osteoblast-like cells.Methods: In this in vitro study, zinc oxide-eugenol (ZOE), eugenol-free zinc oxide (ZONE), and resin (R) cements prepared in cylindrical forms with a similar dimension were used. Each cement was placed on titanium disks to prepare cemented titanium samples. MTT assay was applied at 24- and 72-hour, and 7-day intervals to evaluate cytotoxicity.Results: All the cements decreased cell viability in both cell lines significantly. None of the cements exhibited cellular viability percentages higher than the minimum percentage (70%) required for biocompatibility. The cemented titaniu...
Abstract Context Bacillus anthracis secretes a tripartite toxin comprising protective antigen (PA... more Abstract Context Bacillus anthracis secretes a tripartite toxin comprising protective antigen (PA), edema factor (EF), and lethal factor (LF). The human anthrax vaccine is mainly composed of the anthrax protective antigen (PA). Considerable efforts are being directed towards improving the efficacy of vaccines because the use of commercial anthrax vaccines (human/veterinary) is associated with several limitations. Objective In this study, a triple chimeric antigen referred to as ELP (gene accession no: MT590758) comprising highly immunogenic domains of PA, LF, and EF was designed, constructed, and assessed for the immunization capacity against anthrax in a guinea pig model. Materials and methods Immunization was carried out considering antigen titration and immunization protocol. The immunoprotective efficacy of the ELP was evaluated in guinea pigs and compared with the potency of veterinary anthrax vaccine using a challenge test with B. anthracis 17JB strain spores. Results The results demonstrated that the ELP antigen induced strong humoral responses. The T-cell response of the ELP was found to be similar to PA, and showed that the ELP could protect 100%, 100%, 100%, 80% and 60% of the animals from 50, 70, 90, 100 and 120 times the minimum lethal dose (MLD, equal 5 × 105 spore/ml), respectively, which killed control animals within 48 h. Discussion and conclusions It is concluded that the ELP antigen has the necessary requirement for proper immunization against anthrax and it can be used to develop an effective recombinant vaccine candidate against anthrax.
Abstract Collagen as a biomaterial is commonly obtained from terrestrial animals. However, nowada... more Abstract Collagen as a biomaterial is commonly obtained from terrestrial animals. However, nowadays, the use of collagen with terrestrial animals' sources due to possible transmission of infectious diseases and religious beliefs is restricted. This study was conducted to evaluate the physicochemical characterization, morphology, and biocompatibility of extracted collagen from silver carp fish skin by-product. Type I collagen was extracted from the silver carp fish skin by-product using acetic acid and pepsin enzyme. The results showed that the extraction yield of ASC and PSC was 42.85% and 58.75% (on a dry weight basis), respectively. The presence of two different α-chains confirmed the type I collagen structure for both collagens. Moreover, FTIR spectra investigation demonstrated the triple helical in ASC and PSC collagens. The PSC showed higher imino acids content than ASC. Hence, the fractional viscosity and DSC curves revealed higher Td (30 °C), Ts (81 °C), and Tm (209 °C) for PSC than ASC (28.73 °C, 77 °C, and 187 °C, respectively). Finally, observations of the microscopic and the cell viability evaluation confirmed biocompatibility and suitable structure to cell growth. Accordingly, the obtained collagen from silver carp fish skin can be a proper alternative for terrestrial animals’ collagen and a safe biomaterial for biomedical use.
Abstract In the present study, Poly l -lactic acid (PLLA) resin compatible with digital light pro... more Abstract In the present study, Poly l -lactic acid (PLLA) resin compatible with digital light processing (DLP) 3D printing method was synthesized to produce hard tissue scaffolds. PLLA has been chosen as a decent material to mimic biological structures due to its relatively high strength as well as proper biocompatibility and biodegradation rate. After synthesis and functionalization of PLLA, using a facile method, porous models with 600-micron pore size and 70 % nominal porosity were designed and fabricated via DLP technique in order to investigate the effects of the two process parameters, light exposure time and dye concentration, on compressive strength and morphological features of the printed samples. The experimental results were then reconciled with plotted working curves for each dye concentration to validate the defined exposure time levels. It was concluded that the synthesized polymer and the used method of 3D printing are suitable for fabricating scaffolds with intricate structures. Moreover, by conducting the compression test, a maximum 2.2 MPa strength was achieved for the sample with minimum dye concentration and maximum exposure time. From the biological point of view, no cytotoxic effect was seen after a 3-day in vitro cell viability testing. Altogether, it was shown that optimal adjustment of the process parameters is essential to achieve appropriate dimensional and mechanical properties, which were acknowledged by plotted working curves.
1St Tabriz International Life Science Conference and 12th Iran Biophysical Chemistry Conference, May 22, 2013
Introduction : The PCR is an indispensable method in biotechnology. One of the limitations of PCR... more Introduction : The PCR is an indispensable method in biotechnology. One of the limitations of PCR is the size of the amplified DNA product. The basis for this constraint is the error rate and low proccessivity and elongation rate of different thermostable DNA polymerases. However, if an efficient thermostable PCR enzyme with higher accuracy, elongation velocity and proccessivity such as KOD DNA polymerase is used an increased yield of a high fidelity long_range PCR products can be produced. The aim of this study was to optimize of the production and purification of a functional KOD DNA polymerase. Method : We have purchased a pET17b plasmid that had KOD DNA polymerase gene in multiple cloning site and transform it in the expressing strain BL21(DE3) containing the pLysE or pLysS plasmid. Production of the 90-kDa protein was induced by different concentrations of IPTG. Purification were performed by heating, ammonium sulfate precipitation and chromatography with DEAE-sepharose. Purified protein was identified by SDS-PAGE and AgNO3 staining. The activity of the obtained enzyme was measured by comparing the intensities of the produced DNA bands in PCR reactions. Results: The correct framing of the gene and its orientation were analyzed by digestion and colony PCR. The plasmid was stable in BL21(DE3) containing the pLysS plasmid. Addition of glycerol, Triton X-100, Tween 20, dextrose and protease inhibitor cocktail increased enzyme stability. Conclusions: We have found that pET17b expression vector is toxic or unstable in the expressing strain BL21(DE3), even in the absence of induction. However BL21(DE3) containing the pLysS plasmid suppresses expression prior to induction and was able to produce greater amount of enzyme after induction by IPTG.
Artificial cells, nanomedicine, and biotechnology, Jan 26, 2018
Zein nanoparticles as a carrier system for BMP6-derived peptide were prepared by liquid-liquid ph... more Zein nanoparticles as a carrier system for BMP6-derived peptide were prepared by liquid-liquid phase separation procedure and characterized with SEM, DLS, FTIR and thermogravimetric methods. After peptide encapsulation, nanoparticle size increased from 236.3 ± 92.2 nm to 379.4 ± 116.8 nm. The encapsulation efficiency of peptide was 72.6% and the release of peptide from Zein nanoparticles was partly sustained in trypsin containing phosphate buffered saline (pH 7.4) for up to 14 days. Peptide-loaded nanoparticles showed similar cell viability compared with blank ones. ALP activity of C2C12 cells treated with peptide-loaded nanoparticles (500 µg/mL) was evaluated 7, 14, 21 and 28 days after culture. In peptide-loaded nanoparticles, ALP activity was significantly higher (p < .05) compared with other groups at day 14. Alizarin Red S staining showed, C2C12 cells behind peptide-loaded nanoparticles had significantly (p < .05) higher calcium deposition at day 21. The results of RT-qPC...
The design of a new truncated and engineered alpha1-antitrypsin based on theoretical studies: an ... more The design of a new truncated and engineered alpha1-antitrypsin based on theoretical studies: an antiprotease therapeutics for pulmonary diseases
Cholesteryl ester transfer protein (CETP) catalysis the exchange of triglycerides (TG) and choles... more Cholesteryl ester transfer protein (CETP) catalysis the exchange of triglycerides (TG) and cholesterol esters among plasma lipoproteins. It has been shown that variations at CETP locus is important in the levels and activity of CETP and high density lipoprotein (HDL) plasma concentration. In this research, we assessed the relationship between TaqIB CETP polymorphism and high density lipoprotein-cholestrol (HDL-C) concentration in a study sample of 128 Iranian residents. 1Based on our investigations, it was shown that presence of B1B1 genotype related to TaqIB polymorphism was negatively correlated with HDL-C and HDL2-C, but positively correlated with LDL-C. On the other hand, no correlation with HDL3-C was observed. Analysis of HDL-C subfraction in CHD and control subjects revealed that variation of the CETP TaqI B locus was significantly associated with concentration of HDL2-C subclass. Homozygous and heterozygous carried the B2 alleles, compared with B1 homozygote had significantl...
Human epidermal growth factor receptor 2 (HER2)-positive breast cancer represents approximately 1... more Human epidermal growth factor receptor 2 (HER2)-positive breast cancer represents approximately 15-30% of all invasive breast cancers. Despite the recent advances in therapeutic practices of HER2 subtype, drug resistance and tumor recurrence still have remained as major problems. Drug discovery is a long and difficult process, so the aim of this study is to find potential new application for existing therapeutic agents. Gene expression data for breast invasive carcinoma were retrieved from The Cancer Genome Atlas (TCGA) database. The normal and tumor samples were analyzed using Linear Models for Microarray Data (LIMMA) R package in order to find the differentially expressed genes (DEGs). These genes were used as entry for the library of integrated network-based cellular signatures (LINCS) L1000CDS2 software and suggested 24 repurposed drugs. According to the obtained results, some of these drugs including vorinostat, mocetinostat, alvocidib, CGP-60474, BMS-387032, AT-7519, and curcumin have significant functional similarity and structural correlation with FDA-approved breast cancer drugs. Based on the drug-target network, which consisted of the repurposed drugs and their target genes, the aforementioned drugs had the highest degrees. Moreover, the experimental approach verified curcumin as an effective therapeutic agent for HER2 positive breast cancer. Hence, our work suggested that some repurposed drugs based on gene expression data can be noticed as potential drugs for the treatment of HER2-positive breast cancer.
Due to the unique features of xenografts including large supply from donors, minimal risk of huma... more Due to the unique features of xenografts including large supply from donors, minimal risk of human disease transmission, and the lower cost of preparation and production compared to autografts and allografts, they are considered as attractive alternatives to traditional bone grafts. The animal source accessibility and production process have a direct correlation with the cost and quality of the final product. To evaluate whether the animal source of the bone has any effect on the physicochemical and histological properties of the final xenograft, three deproteinized bone grafts were prepared from sources that are easily available in Iran, including the bovine (DBB), camel (DCB), and ostrich (DOB).
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