Hamid Nazarian | Shahid Beheshti University of Medical Sciences (original) (raw)

Papers by Hamid Nazarian

The International journal of artificial organs, 2010

In this study, PLGA microspheres were prepared using a water-in-oil-in-water emulsion/solvent eva... more In this study, PLGA microspheres were prepared using a water-in-oil-in-water emulsion/solvent evaporation technique. Some microspheres were coated with poly-L-lysine (an extracellular matrix (ECM) component), and then pluripotent P19 embryonic carcinoma cells were seeded on them. P19 cells attached onto the PLGA microspheres; subsequently, by adding retinoic acid (RA) to cell culture medium as a neurogenic inducer (RA was released from the microspheres), the cells differentiated into neural cells. Size and morphology of PLGA microspheres was characterized by scanning electron microscopy (SEM). Neurogenic differentiation was studied by immunofluorescent staining, real-time polymerase chain reaction (RT-PCR), and light microscopy. Histological assay showed that more cells attached onto microspheres coated with poly-L-lysine than the uncoated group. Immunofluoresent staining and RT-PCR analysis for ß-Tubulin, Nestin and Pax6 genes indicated differentiation of P19 cells into neural cells on both coated and uncoated microspheres. It was found that a high surface area of microspheres improves cell attachment and expansion, which was significantly increased in those coated with poly-L-lysine. Finally, these results highlight the versatility of these sample scaffolds as a model system for nerve tissue engineering.

The Journal of oral implantology, 2013

This study aimed to assess vertical bone augmentation with simultaneous implant placement in rabb... more This study aimed to assess vertical bone augmentation with simultaneous implant placement in rabbit tibiae using particulate mineralized bone/fibrin glue/mesenchymal stem cell. Bone marrow was aspirated from tibiae of five 10-week-old New Zealand White male rabbits. Right and left tibiae of each rabbit were prepared, and a 3-mm protruding implant from tibial bone was placed in each side. Particulate allogenic bone/fibrin glue/mesenchymal stem cell combination was placed around test implants and particulate bone graft/fibrin glue around controls. Two months postoperatively, the animals were euthanized, and sections were prepared for histological analysis. The mean amount of vertical bone length was higher in the experimental group than the control group (2.09 mm vs 1.03 mm; P < .05). New supracrestal trabecular bone formation was also significantly higher in the test group (28.5 ± 4.5% vs 4.3 ± 1.8%; P < .05). Mesenchymal stem cell/particulate allograft/fibrin glue appears to be a promising combination for vertical bone augmentation around simultaneously inserted implants in rabbit tibia.

The Journal of oral implantology, 2012

The Journal of oral implantology, 2013

Journal of biomedical materials research. Part B, Applied biomaterials, 2014

Hydroxyapatite with different characteristics in terms of morphology and chemistry were prepared ... more Hydroxyapatite with different characteristics in terms of morphology and chemistry were prepared via conventional sintering and low temperature biomimetic mineralization methods. The biomineralization route introduced nanocrystalline carbonate-substituted hydroxyapatite (n-CHA) with needle-like crystals ranging 20–30 nm whereas sintered HA (S-HA) comprised of polygonal grains ranging 2–5 μm. The response of fibroblastic cells was investigated using the extract of the samples whereas Wistar rat-derived mesenchymal stem cells (MSCs) were evaluated on top of each sample while maintaining in an osteogenic-free medium. The proliferation, activity, and morphology of adherent MSCs were determined at different culturing periods. The osteogenic differentiation of MSCs was also assayed by determining expression of runx2, osteonectin, osteopontin, and osteocalcin genes using real time-PCR analysis. The fibroblastic cells exhibited better proliferation rate at the presence of n-CHA compared to S-HA. Furthermore, the MSCs attached and spread well on both n-CHA and S-HA with better proliferation rate and alkaline phosphatase activity on n-CHA. Interestingly, the osteogenic differentiation of MSCs on n-CHA was confirmed by the expression of bone specific proteins whereas poor expression of these proteins was detected for the cells on S-HA. The results showed that the role of morphology, crystallinity, and chemistry of hydroxyapatite is crucial for osteogenesis differentiation of MSCs. The results predict osteoinductivity of n-CHA, because MSCs differentiation occurred at the absence of osteogenic medium. However, in vivo data are also required to support this suggestion. © 2013 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater, 102B: 108–118, 2014.

Journal of endodontics, 2014

Mineral trioxide aggregate (MTA) and calcium-enriched mixture (CEM) have shown osteogenic/cemento... more Mineral trioxide aggregate (MTA) and calcium-enriched mixture (CEM) have shown osteogenic/cementogenic/dentinogenic activities; however, their mechanism of action is not fully understood. We aimed to evaluate the effect of these biomaterials on odontogenic differentiation of human dental pulp stem cells (DPSCs). Flow cytometry with stem cell markers for the confirmation of stemness and homogeneity was first performed. Then isolated DPSCs were seeded on prepared discs of MTA, CEM, differentiation medium (DM), and growth medium (GM) and incubated up to 14 days. Concentrations of transforming growth factor-β1, bone morphogenetic protein (BMP)2, BMP4, and fibroblast growth factor 4 were measured at each interval using an enzyme-linked immunosorbent assay reader. Gene expression of dentin sialophosphoprotein, dentin matrix protein 1, and the cytokines were evaluated by reverse-transcription polymerase chain reaction. To evaluate the cell morphology, scanning electron micrographs were taken; mineralization potential was evaluated using alizarin red S staining. Scanning electron micrographs showed that DPSCs spread/adhered/proliferated similarly on MTA and CEM. On day 14, alizarin red S staining confirmed that mineralization occurred in all groups except GM. Expressions of dentin matrix protein 1 and dentin sialophosphoprotein genes were similar in the CEM, MTA, and DM groups; they were significantly higher compared with the GM group (P < .05). A greater amount of transforming growth factor-β1 gene was expressed in MTA compared with the other groups (P < .05). However, the expression of fibroblast growth factor 4 and BMP2 genes was significantly greater in the CEM group (P < .05). In all the tested groups, the expression of BMP4 was less than GM (P < .01); however, CEM and DM were similar but more than MTA (P < .05). Concentrations of protein product detected using an enzyme-linked immunosorbent assay reader confirmed these gene expressions. MTA and CEM can induce osteo-/odontogenic-like phenotype differentiation of human DPSCs; however, they stimulate different gene expressions and growth factor release.

Molecular biology reports, 2014

The combination of bioceramics and stem cells has attracted the interest of research community fo... more The combination of bioceramics and stem cells has attracted the interest of research community for bone tissue engineering applications. In the present study, a combination of Bio-Oss® and type 1 collagen gel as scaffold were loaded with human adipose-tissue derived mesenchymal stem cells (AT-MSCs) after isolation and characterization, and the capacity of them for bone regeneration was investigated in rat critical size defects using digital mammography, multi-slice spiral computed tomography imaging and histological analysis. 8 weeks after implantation, no mortality or sign of inflammation was observed in the site of defect. According to the results of imaging analysis, a higher level of bone regeneration was observed in the rats receiving Bio-Oss®-Gel compared to untreated group. In addition, MSC-seeded Bio-Oss-Gel induced the highest bone reconstruction among all groups. Histological staining confirmed these findings and impressive osseointegration was observed in MSC-seeded Bio-Oss-Gel compared with Bio-Oss-Gel. On the whole, it was demonstrated that combination of AT-MSCs, Bio-Oss and Gel synergistically enhanced bone regeneration and reconstruction and also could serve as an appropriate structure to bone regenerative medicine and tissue engineering application.

Iranian journal of reproductive medicine, 2014

The Wnt/β-The Wnt/β-catenin signaling pathway is involved in many developmental processes in both... more The Wnt/β-The Wnt/β-catenin signaling pathway is involved in many developmental processes in both fetal and adult life; its abnormalities can lead to disorders including several types of cancers and malfunction of specific cells and tissues in both animals and humans. Its role in reproductive processes has been proven. Objective: This study was designed to evaluate the expression of the key regulator of this signaling pathway GSK3-β and its presumed role in azoospermia. Materials and Methods: WNT3a protein concentration and GSK3-β gene expression levels were measured and compared between two groups of infertile men. The test groups consisted of 10 patients with obstructive and 10 nonobstructive azoospermia. The control group was selected among healthy men after vasectomies that were willing to conceive a child using a testicular biopsy technique. Samples were obtained by testicular biopsy and screened for the most common mutations (84, 86 and 255) in the SRY region before analyzing. GSK3-β gene expression was assessed quantitatively by real time-PCR. Results: The WNT3a protein concentration had no significant difference between the two test groups and controls. Expression of GSK3-β was down-regulated in nonobstructive azoospermia (3.10±0.19) compared with normal (7.12±0.39) and obstructive azoospermia (6.32±0.42) groups (p=0.001). Conclusion: Down-regulation of GSK-3β may cause to non-obstructive azoospermia. Regulation and modification of GSK-3β gene expression by drugs could be used as a therapeutic solution.

Advances in Applied Ceramics, 2009

In the present work, calcium phosphate cement was made by mixing a solid phase and blood plasma a... more In the present work, calcium phosphate cement was made by mixing a solid phase and blood plasma as liquid phase. The basic properties of the cement (called BPC) were compared with those of conventional calcium phosphate cement (c-CPC) where distilled water was used as liquid. BPC had better consistency and injectability than c-CPC but longer setting time. In both cements, the reactants were converted into apatite phase after immersing in simulated body fluid but the phase formed in BPC had lower crystallinity than the phase formed in c-CPC. The set BPC was stronger than c-CPC, having a compressive strength (CS) of about 2-6 MPa after 24 h incubation at 37uC. The CS reduced during soaking at early stage but was relatively improved at the end of soaking period (day 7). In contrast, an increase in CS was observed in c-CPC during soaking period.

Materials Science and Engineering: C, 2011

In this study, different concentrations of Li2O (0–12wt.%) were substituted for Na2O in 45S5 biog... more In this study, different concentrations of Li2O (0–12wt.%) were substituted for Na2O in 45S5 bioglass® and the effect of these substitutions on both in vitro apatite formation ability and osteoblastic cell responses was studied. For these purposes, the structural and topographical properties of the glasses were studied using, XRD, FTIR, SEM/EDXA and AFM techniques, before and after storing in simulated

Yakhteh Medical Journal, Jan 1, 2008

Journal of Materials …, Jan 1, 2010

Colloids and Surfaces B: …, Jan 1, 2009

Journal of Biomedical …, Jan 1, 2009

In the present study, series of Zn incorporated calcium sulfate bone cements, with different amou... more In the present study, series of Zn incorporated calcium sulfate bone cements, with different amounts of doped Zn(0, 0.74, 1.97, 3.05, 4.21 wt %) were prepared by mixing a calcium sulfate hemihydrate powder and solutions of zinc sulfate, and the effect of zinc-doping on some physical, physico-chemical, and biological properties of the cements were investigated. Pure calcium sulfate cement was also made as control, with the mentioned powder and distilled water as liquid phase. The initial setting time and compressive strength of the cement significantly changed from 17 min and 3.2 MPa for the pure calcium sulfate to 6 min and 6 MPa for the Zn-added calcium sulfate, respectively. Compared to pure calcium sulfate, more gypsum precipitates were formed in the zinc sulfate added samples with a morphology of thin, elongated, and rod-shaped crystals. The biological properties of the samples were analyzed in the terms of cell viability and cell activity on human osteosarcoma (G-292) using MTT assay and alkaline phosphatase (ALP) activity in the cell culture medium. The best increased cell density and ALP activity were achieved for the calcium sulfate cement with a content of 0.74 wt % Zn, whereas a toxic behavior was observed for the samples with Zn concentrations more than 1.97%.

Iranian Journal of Basic …

... Search result page. Title: Ex vivo Expansion and Differentiation of Mesenchymal Stem Cells fr... more ... Search result page. Title: Ex vivo Expansion and Differentiation of Mesenchymal Stem Cells from Goat Bone Marrow. Author: Mohamadreza Baghaban Eslaminejad ; Hamid Nazarian ; Fahimeh Falahi ; Leila Taghiyar ; Mohamad Taghi Daneshzadeh. ...

Oral Surgery, Oral Medicine, …, Jan 1, 2008

The International journal of artificial organs, 2010

In this study, PLGA microspheres were prepared using a water-in-oil-in-water emulsion/solvent eva... more In this study, PLGA microspheres were prepared using a water-in-oil-in-water emulsion/solvent evaporation technique. Some microspheres were coated with poly-L-lysine (an extracellular matrix (ECM) component), and then pluripotent P19 embryonic carcinoma cells were seeded on them. P19 cells attached onto the PLGA microspheres; subsequently, by adding retinoic acid (RA) to cell culture medium as a neurogenic inducer (RA was released from the microspheres), the cells differentiated into neural cells. Size and morphology of PLGA microspheres was characterized by scanning electron microscopy (SEM). Neurogenic differentiation was studied by immunofluorescent staining, real-time polymerase chain reaction (RT-PCR), and light microscopy. Histological assay showed that more cells attached onto microspheres coated with poly-L-lysine than the uncoated group. Immunofluoresent staining and RT-PCR analysis for ß-Tubulin, Nestin and Pax6 genes indicated differentiation of P19 cells into neural cells on both coated and uncoated microspheres. It was found that a high surface area of microspheres improves cell attachment and expansion, which was significantly increased in those coated with poly-L-lysine. Finally, these results highlight the versatility of these sample scaffolds as a model system for nerve tissue engineering.

The Journal of oral implantology, 2013

This study aimed to assess vertical bone augmentation with simultaneous implant placement in rabb... more This study aimed to assess vertical bone augmentation with simultaneous implant placement in rabbit tibiae using particulate mineralized bone/fibrin glue/mesenchymal stem cell. Bone marrow was aspirated from tibiae of five 10-week-old New Zealand White male rabbits. Right and left tibiae of each rabbit were prepared, and a 3-mm protruding implant from tibial bone was placed in each side. Particulate allogenic bone/fibrin glue/mesenchymal stem cell combination was placed around test implants and particulate bone graft/fibrin glue around controls. Two months postoperatively, the animals were euthanized, and sections were prepared for histological analysis. The mean amount of vertical bone length was higher in the experimental group than the control group (2.09 mm vs 1.03 mm; P < .05). New supracrestal trabecular bone formation was also significantly higher in the test group (28.5 ± 4.5% vs 4.3 ± 1.8%; P < .05). Mesenchymal stem cell/particulate allograft/fibrin glue appears to be a promising combination for vertical bone augmentation around simultaneously inserted implants in rabbit tibia.

The Journal of oral implantology, 2012

The Journal of oral implantology, 2013

Journal of biomedical materials research. Part B, Applied biomaterials, 2014

Hydroxyapatite with different characteristics in terms of morphology and chemistry were prepared ... more Hydroxyapatite with different characteristics in terms of morphology and chemistry were prepared via conventional sintering and low temperature biomimetic mineralization methods. The biomineralization route introduced nanocrystalline carbonate-substituted hydroxyapatite (n-CHA) with needle-like crystals ranging 20–30 nm whereas sintered HA (S-HA) comprised of polygonal grains ranging 2–5 μm. The response of fibroblastic cells was investigated using the extract of the samples whereas Wistar rat-derived mesenchymal stem cells (MSCs) were evaluated on top of each sample while maintaining in an osteogenic-free medium. The proliferation, activity, and morphology of adherent MSCs were determined at different culturing periods. The osteogenic differentiation of MSCs was also assayed by determining expression of runx2, osteonectin, osteopontin, and osteocalcin genes using real time-PCR analysis. The fibroblastic cells exhibited better proliferation rate at the presence of n-CHA compared to S-HA. Furthermore, the MSCs attached and spread well on both n-CHA and S-HA with better proliferation rate and alkaline phosphatase activity on n-CHA. Interestingly, the osteogenic differentiation of MSCs on n-CHA was confirmed by the expression of bone specific proteins whereas poor expression of these proteins was detected for the cells on S-HA. The results showed that the role of morphology, crystallinity, and chemistry of hydroxyapatite is crucial for osteogenesis differentiation of MSCs. The results predict osteoinductivity of n-CHA, because MSCs differentiation occurred at the absence of osteogenic medium. However, in vivo data are also required to support this suggestion. © 2013 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater, 102B: 108–118, 2014.

Journal of endodontics, 2014

Mineral trioxide aggregate (MTA) and calcium-enriched mixture (CEM) have shown osteogenic/cemento... more Mineral trioxide aggregate (MTA) and calcium-enriched mixture (CEM) have shown osteogenic/cementogenic/dentinogenic activities; however, their mechanism of action is not fully understood. We aimed to evaluate the effect of these biomaterials on odontogenic differentiation of human dental pulp stem cells (DPSCs). Flow cytometry with stem cell markers for the confirmation of stemness and homogeneity was first performed. Then isolated DPSCs were seeded on prepared discs of MTA, CEM, differentiation medium (DM), and growth medium (GM) and incubated up to 14 days. Concentrations of transforming growth factor-β1, bone morphogenetic protein (BMP)2, BMP4, and fibroblast growth factor 4 were measured at each interval using an enzyme-linked immunosorbent assay reader. Gene expression of dentin sialophosphoprotein, dentin matrix protein 1, and the cytokines were evaluated by reverse-transcription polymerase chain reaction. To evaluate the cell morphology, scanning electron micrographs were taken; mineralization potential was evaluated using alizarin red S staining. Scanning electron micrographs showed that DPSCs spread/adhered/proliferated similarly on MTA and CEM. On day 14, alizarin red S staining confirmed that mineralization occurred in all groups except GM. Expressions of dentin matrix protein 1 and dentin sialophosphoprotein genes were similar in the CEM, MTA, and DM groups; they were significantly higher compared with the GM group (P < .05). A greater amount of transforming growth factor-β1 gene was expressed in MTA compared with the other groups (P < .05). However, the expression of fibroblast growth factor 4 and BMP2 genes was significantly greater in the CEM group (P < .05). In all the tested groups, the expression of BMP4 was less than GM (P < .01); however, CEM and DM were similar but more than MTA (P < .05). Concentrations of protein product detected using an enzyme-linked immunosorbent assay reader confirmed these gene expressions. MTA and CEM can induce osteo-/odontogenic-like phenotype differentiation of human DPSCs; however, they stimulate different gene expressions and growth factor release.

Molecular biology reports, 2014

The combination of bioceramics and stem cells has attracted the interest of research community fo... more The combination of bioceramics and stem cells has attracted the interest of research community for bone tissue engineering applications. In the present study, a combination of Bio-Oss® and type 1 collagen gel as scaffold were loaded with human adipose-tissue derived mesenchymal stem cells (AT-MSCs) after isolation and characterization, and the capacity of them for bone regeneration was investigated in rat critical size defects using digital mammography, multi-slice spiral computed tomography imaging and histological analysis. 8 weeks after implantation, no mortality or sign of inflammation was observed in the site of defect. According to the results of imaging analysis, a higher level of bone regeneration was observed in the rats receiving Bio-Oss®-Gel compared to untreated group. In addition, MSC-seeded Bio-Oss-Gel induced the highest bone reconstruction among all groups. Histological staining confirmed these findings and impressive osseointegration was observed in MSC-seeded Bio-Oss-Gel compared with Bio-Oss-Gel. On the whole, it was demonstrated that combination of AT-MSCs, Bio-Oss and Gel synergistically enhanced bone regeneration and reconstruction and also could serve as an appropriate structure to bone regenerative medicine and tissue engineering application.

Iranian journal of reproductive medicine, 2014

The Wnt/β-The Wnt/β-catenin signaling pathway is involved in many developmental processes in both... more The Wnt/β-The Wnt/β-catenin signaling pathway is involved in many developmental processes in both fetal and adult life; its abnormalities can lead to disorders including several types of cancers and malfunction of specific cells and tissues in both animals and humans. Its role in reproductive processes has been proven. Objective: This study was designed to evaluate the expression of the key regulator of this signaling pathway GSK3-β and its presumed role in azoospermia. Materials and Methods: WNT3a protein concentration and GSK3-β gene expression levels were measured and compared between two groups of infertile men. The test groups consisted of 10 patients with obstructive and 10 nonobstructive azoospermia. The control group was selected among healthy men after vasectomies that were willing to conceive a child using a testicular biopsy technique. Samples were obtained by testicular biopsy and screened for the most common mutations (84, 86 and 255) in the SRY region before analyzing. GSK3-β gene expression was assessed quantitatively by real time-PCR. Results: The WNT3a protein concentration had no significant difference between the two test groups and controls. Expression of GSK3-β was down-regulated in nonobstructive azoospermia (3.10±0.19) compared with normal (7.12±0.39) and obstructive azoospermia (6.32±0.42) groups (p=0.001). Conclusion: Down-regulation of GSK-3β may cause to non-obstructive azoospermia. Regulation and modification of GSK-3β gene expression by drugs could be used as a therapeutic solution.

Advances in Applied Ceramics, 2009

In the present work, calcium phosphate cement was made by mixing a solid phase and blood plasma a... more In the present work, calcium phosphate cement was made by mixing a solid phase and blood plasma as liquid phase. The basic properties of the cement (called BPC) were compared with those of conventional calcium phosphate cement (c-CPC) where distilled water was used as liquid. BPC had better consistency and injectability than c-CPC but longer setting time. In both cements, the reactants were converted into apatite phase after immersing in simulated body fluid but the phase formed in BPC had lower crystallinity than the phase formed in c-CPC. The set BPC was stronger than c-CPC, having a compressive strength (CS) of about 2-6 MPa after 24 h incubation at 37uC. The CS reduced during soaking at early stage but was relatively improved at the end of soaking period (day 7). In contrast, an increase in CS was observed in c-CPC during soaking period.

Materials Science and Engineering: C, 2011

In this study, different concentrations of Li2O (0–12wt.%) were substituted for Na2O in 45S5 biog... more In this study, different concentrations of Li2O (0–12wt.%) were substituted for Na2O in 45S5 bioglass® and the effect of these substitutions on both in vitro apatite formation ability and osteoblastic cell responses was studied. For these purposes, the structural and topographical properties of the glasses were studied using, XRD, FTIR, SEM/EDXA and AFM techniques, before and after storing in simulated

Yakhteh Medical Journal, Jan 1, 2008

Journal of Materials …, Jan 1, 2010

Colloids and Surfaces B: …, Jan 1, 2009

Journal of Biomedical …, Jan 1, 2009

In the present study, series of Zn incorporated calcium sulfate bone cements, with different amou... more In the present study, series of Zn incorporated calcium sulfate bone cements, with different amounts of doped Zn(0, 0.74, 1.97, 3.05, 4.21 wt %) were prepared by mixing a calcium sulfate hemihydrate powder and solutions of zinc sulfate, and the effect of zinc-doping on some physical, physico-chemical, and biological properties of the cements were investigated. Pure calcium sulfate cement was also made as control, with the mentioned powder and distilled water as liquid phase. The initial setting time and compressive strength of the cement significantly changed from 17 min and 3.2 MPa for the pure calcium sulfate to 6 min and 6 MPa for the Zn-added calcium sulfate, respectively. Compared to pure calcium sulfate, more gypsum precipitates were formed in the zinc sulfate added samples with a morphology of thin, elongated, and rod-shaped crystals. The biological properties of the samples were analyzed in the terms of cell viability and cell activity on human osteosarcoma (G-292) using MTT assay and alkaline phosphatase (ALP) activity in the cell culture medium. The best increased cell density and ALP activity were achieved for the calcium sulfate cement with a content of 0.74 wt % Zn, whereas a toxic behavior was observed for the samples with Zn concentrations more than 1.97%.

Iranian Journal of Basic …

... Search result page. Title: Ex vivo Expansion and Differentiation of Mesenchymal Stem Cells fr... more ... Search result page. Title: Ex vivo Expansion and Differentiation of Mesenchymal Stem Cells from Goat Bone Marrow. Author: Mohamadreza Baghaban Eslaminejad ; Hamid Nazarian ; Fahimeh Falahi ; Leila Taghiyar ; Mohamad Taghi Daneshzadeh. ...

Oral Surgery, Oral Medicine, …, Jan 1, 2008