Zhongbai Zhang, Xueting Qin, Jingxun Chen, Yanchun Li, Huaxin Chen, H. Xie, Min Yang, Chuang Li, Zhenghui Wang, Mei Zhang
Coronary atherosclerotic heart disease is an important threat to human health. The pathological basis is atherosclerosis, and foam cell formation is the key link in the initiation of atherosclerosis. Here, foam cell models were established using 50 ng/ml oxidized low-density lipoprotein (ox-LDL) to stimulate in vitro cultures of THP-1 cells for 72 h. The expression of ZNF580, a Cys2–His2 (C2H2) zinc finger protein containing 172 amino acids that was originally cloned by screening a human aortic cDNA library, was measured in foam cells, and its interaction with various regulatory factors during foam cell formation was investigated. Oil red O (ORO) staining was used to observe cell morphology and intracellular lipid levels. Lentivirus transfection was used to induce high ZNF580 expression (Ad-ZNF580) and low ZNF580 expression (Si-ZNF580) in THP-1 cells, and a fluorescent inverted microscope was used to observe the distribution of ZNF580 immunofluorescence to deduce the transfection rate. RNA and total protein were extracted, and the expression levels of ZNF580, cluster of differentiation 36 (CD36), peroxisome proliferator activated receptor-γ (PPAR-γ), ATP-binding cassette transporter A1 (ABCA1) and apolipoprotein E (ApoE) were measured by real-time quantitative PCR. The protein levels were examined by western blotting to evaluate the interaction between ZNF580 and associated regulatory factors. ZNF580 can significantly increase the expression levels of ApoE and ABCA1 and significantly decrease the expression levels of CD36 and PPAR-γ, suggesting that ZNF580-mediated inhibition of foam cell formation is associated with the PPAR-γ-CD36 signalling pathway. Based on these findings, ZNF580 might be a potential therapeutic candidate for the treatment of coronary atherosclerotic heart disease.
{"title":"The role and mechanism of the zinc finger gene ZNF580 in foam cell formation","authors":"Zhongbai Zhang, Xueting Qin, Jingxun Chen, Yanchun Li, Huaxin Chen, H. Xie, Min Yang, Chuang Li, Zhenghui Wang, Mei Zhang","doi":"10.3233/jcb-220063","DOIUrl":"https://doi.org/10.3233/jcb-220063","url":null,"abstract":"Coronary atherosclerotic heart disease is an important threat to human health. The pathological basis is atherosclerosis, and foam cell formation is the key link in the initiation of atherosclerosis. Here, foam cell models were established using 50 ng/ml oxidized low-density lipoprotein (ox-LDL) to stimulate in vitro cultures of THP-1 cells for 72 h. The expression of ZNF580, a Cys2–His2 (C2H2) zinc finger protein containing 172 amino acids that was originally cloned by screening a human aortic cDNA library, was measured in foam cells, and its interaction with various regulatory factors during foam cell formation was investigated. Oil red O (ORO) staining was used to observe cell morphology and intracellular lipid levels. Lentivirus transfection was used to induce high ZNF580 expression (Ad-ZNF580) and low ZNF580 expression (Si-ZNF580) in THP-1 cells, and a fluorescent inverted microscope was used to observe the distribution of ZNF580 immunofluorescence to deduce the transfection rate. RNA and total protein were extracted, and the expression levels of ZNF580, cluster of differentiation 36 (CD36), peroxisome proliferator activated receptor-γ (PPAR-γ), ATP-binding cassette transporter A1 (ABCA1) and apolipoprotein E (ApoE) were measured by real-time quantitative PCR. The protein levels were examined by western blotting to evaluate the interaction between ZNF580 and associated regulatory factors. ZNF580 can significantly increase the expression levels of ApoE and ABCA1 and significantly decrease the expression levels of CD36 and PPAR-γ, suggesting that ZNF580-mediated inhibition of foam cell formation is associated with the PPAR-γ-CD36 signalling pathway. Based on these findings, ZNF580 might be a potential therapeutic candidate for the treatment of coronary atherosclerotic heart disease.","PeriodicalId":15286,"journal":{"name":"Journal of Cellular Biotechnology","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2022-11-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"46250033","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
S. Joshi, G. Pendyala, Neeta S. Padmawar, Viddyasagar Mopagar
Enamel Matrix Derivatives (EMD) is a novel biomaterial that has been discovered in the late ’90 s and has found numerous uses in the field of periodontics. It is mainly used in areas that require natural regeneration and healing. It has found applications in other branches of dentistry, such as; wound healing, regenerative procedures, endodontics, pedodontics, and others. It mainly consists of amelogenins and other proteins such as amelin, enamelin, tufetelin etc. It is known to increase cementogenesis and improve periodontal attachment. The amount of research done on this product is very little and very scattered. The maximum amount of research has been done regarding its use in periodontic surgery and other related procedures since it had been initially discovered as a material that would increase periodontal attachment. Given its various biological properties, it is safe to say that it has the potential to succeed in other branches. Our review, therefore, aims to collect and present the research performed and ongoing research potential regarding EMD products, which are now available in a gel-based form known as Emdogain.
{"title":"Evidentiary manoeuvrings of neoteric polymer: Emdogain – An Annotation","authors":"S. Joshi, G. Pendyala, Neeta S. Padmawar, Viddyasagar Mopagar","doi":"10.3233/jcb-220078","DOIUrl":"https://doi.org/10.3233/jcb-220078","url":null,"abstract":"Enamel Matrix Derivatives (EMD) is a novel biomaterial that has been discovered in the late ’90 s and has found numerous uses in the field of periodontics. It is mainly used in areas that require natural regeneration and healing. It has found applications in other branches of dentistry, such as; wound healing, regenerative procedures, endodontics, pedodontics, and others. It mainly consists of amelogenins and other proteins such as amelin, enamelin, tufetelin etc. It is known to increase cementogenesis and improve periodontal attachment. The amount of research done on this product is very little and very scattered. The maximum amount of research has been done regarding its use in periodontic surgery and other related procedures since it had been initially discovered as a material that would increase periodontal attachment. Given its various biological properties, it is safe to say that it has the potential to succeed in other branches. Our review, therefore, aims to collect and present the research performed and ongoing research potential regarding EMD products, which are now available in a gel-based form known as Emdogain.","PeriodicalId":15286,"journal":{"name":"Journal of Cellular Biotechnology","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2022-11-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"45684480","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Shivani Sachdeva, Amit Mani, Hiral R. Vora, H. Saluja, Nishant Manka, V. Dehane
Tissue engineering comprises of an array of specialities which combines biology, chemical sciences, engineering and material sciences for the regeneration of diseased tissues. In the novel world of tissue engineering, the fabrication and role of scaffolds is vital. Scaffolds have been engineered in such a fashion that it causes the desirable cellular interactions for the formation of new tissues for medical purposes. Ideal characteristics of scaffold include; three –dimensional and highly porous, should be biocompatible and bioresorbable, should have suitable surface chemistry for cell attachment, proliferation, and differentiation and must have mechanical properties to match those of the tissues at the site of implantation. A high porosity and an adequate pore size are necessary to facilitate cell seeding and diffusion throughout the whole structure of both cells and nutrients. The ever- evolving world of medical science will now in the near future be able to regenerate the lost tissues with the advancements of tissue engineering.
{"title":"Scaffolds in tissue engineering","authors":"Shivani Sachdeva, Amit Mani, Hiral R. Vora, H. Saluja, Nishant Manka, V. Dehane","doi":"10.3233/jcb-220070","DOIUrl":"https://doi.org/10.3233/jcb-220070","url":null,"abstract":"Tissue engineering comprises of an array of specialities which combines biology, chemical sciences, engineering and material sciences for the regeneration of diseased tissues. In the novel world of tissue engineering, the fabrication and role of scaffolds is vital. Scaffolds have been engineered in such a fashion that it causes the desirable cellular interactions for the formation of new tissues for medical purposes. Ideal characteristics of scaffold include; three –dimensional and highly porous, should be biocompatible and bioresorbable, should have suitable surface chemistry for cell attachment, proliferation, and differentiation and must have mechanical properties to match those of the tissues at the site of implantation. A high porosity and an adequate pore size are necessary to facilitate cell seeding and diffusion throughout the whole structure of both cells and nutrients. The ever- evolving world of medical science will now in the near future be able to regenerate the lost tissues with the advancements of tissue engineering.","PeriodicalId":15286,"journal":{"name":"Journal of Cellular Biotechnology","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2022-08-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"46615888","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
G. Pendyala, S. Joshi, A. Mani, Sudhir Dhole, P. Kale
BACKGROUND: Scaling and root planing [SRP] being the mainstay of treatment of periodontitis encompasses unambiguous impediments. antiseptics represent an aid to nonsurgical periodontal therapy. OBJECTIVE: This randomized controlled, split mouth study design with an observation period of three months aims to clinically evaluate the efficacy of ozonised oil and chlorhexidine as an adjunct to SRP. METHODS: Twenty-five patients of both sexes with an age range of 30–65 years diagnosed with chronic periodontitis and having a periodontal probe depth (PD)≥5 mm and CAL≥3 mm on at least 1 site in each quadrant were included in this randomised split mouth design study. Patients were allocated in 2 experimental treatment groups as SRP + chlorhexidine gel (control sites) and with SRP + ozone oil (test sites). The plaque index (PI), gingival index (GI), and periodontal pocket depth (PPD), clinical attachment level (CAL) were recorded at baseline data and after 30 days post-baseline. RESULTS: The present study showed significant results in both the groups with regards to the improvement in the clinical parameters. When comparison was made between the two groups, it has been assessed that the use of the ozonized oil in addition to SRP did not show significant differences when compared to conventional SRP + chlorhexidine. CONCLUSION: For bye to SRP, ozonized oil can be considered as a viable alternative to chlorhexidine in the treatment of periodontitis, especially considering its low toxicity compared to chlorhexidine.
{"title":"The denouement of ozone therapy vying to chlorhexidine in non surgical periodontal therapy","authors":"G. Pendyala, S. Joshi, A. Mani, Sudhir Dhole, P. Kale","doi":"10.3233/jcb-220073","DOIUrl":"https://doi.org/10.3233/jcb-220073","url":null,"abstract":"BACKGROUND: Scaling and root planing [SRP] being the mainstay of treatment of periodontitis encompasses unambiguous impediments. antiseptics represent an aid to nonsurgical periodontal therapy. OBJECTIVE: This randomized controlled, split mouth study design with an observation period of three months aims to clinically evaluate the efficacy of ozonised oil and chlorhexidine as an adjunct to SRP. METHODS: Twenty-five patients of both sexes with an age range of 30–65 years diagnosed with chronic periodontitis and having a periodontal probe depth (PD)≥5 mm and CAL≥3 mm on at least 1 site in each quadrant were included in this randomised split mouth design study. Patients were allocated in 2 experimental treatment groups as SRP + chlorhexidine gel (control sites) and with SRP + ozone oil (test sites). The plaque index (PI), gingival index (GI), and periodontal pocket depth (PPD), clinical attachment level (CAL) were recorded at baseline data and after 30 days post-baseline. RESULTS: The present study showed significant results in both the groups with regards to the improvement in the clinical parameters. When comparison was made between the two groups, it has been assessed that the use of the ozonized oil in addition to SRP did not show significant differences when compared to conventional SRP + chlorhexidine. CONCLUSION: For bye to SRP, ozonized oil can be considered as a viable alternative to chlorhexidine in the treatment of periodontitis, especially considering its low toxicity compared to chlorhexidine.","PeriodicalId":15286,"journal":{"name":"Journal of Cellular Biotechnology","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2022-08-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"44391337","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Cancer is the second most common reason for death in the world. The cancer research over four decades has been reached to the prospective on dysregulation of ions like (Ca2 +, Mg2 +, Na +, K+, or Cl - ) recently. These ions are orchestrated through numerous proteins, ion channels, selectively or non-selectively. However, the dysregulation of these ions and their channel expression are being reported for various diseases but here we have reviewed precisely TRP channels (TRPC and TRPM) for their role in cancer. The transient receptor potential (TRP) channels were first discovered in Drosophila melanogaster in 1989 and since then the superfamily becomes a group of 30 members under six subsections. Interestingly, we found that the TRPC (Canonical) channels, with 6 members, were explored in nine different types of cancers in last two decades. Additionally, we included the TRPM (Melastatin) subfamily and reviewed their role in cancer. Conclusively, these studies support that TRP channel-based therapies must be taken forward for clinical studies. Some channels, such as TRPC6, TRPM7 and TRPM8 were explored extensively in many cancer types which may be a potential target for cancer treatment. However, TRPM8 in lung cancer was reported for reverse association with cell proliferation, which needs to be reverified in lung cancer and other cancers. Besides, some TRPC channels are associated with store-operated calcium entry (SOCE) such as TRPC1, TRPC4 and TRPC6. Interestingly, the TRPC6 role was reported in breast cancer for modulation of Ca2 + through translocation of Orai1 and Orai3.
{"title":"TRPC and TRPM channels: New possible targets for cancer","authors":"Priyanka Verma, R. Rani, Priya T Rao, A. Singh","doi":"10.3233/jcb-220066","DOIUrl":"https://doi.org/10.3233/jcb-220066","url":null,"abstract":"Cancer is the second most common reason for death in the world. The cancer research over four decades has been reached to the prospective on dysregulation of ions like (Ca2 +, Mg2 +, Na +, K+, or Cl - ) recently. These ions are orchestrated through numerous proteins, ion channels, selectively or non-selectively. However, the dysregulation of these ions and their channel expression are being reported for various diseases but here we have reviewed precisely TRP channels (TRPC and TRPM) for their role in cancer. The transient receptor potential (TRP) channels were first discovered in Drosophila melanogaster in 1989 and since then the superfamily becomes a group of 30 members under six subsections. Interestingly, we found that the TRPC (Canonical) channels, with 6 members, were explored in nine different types of cancers in last two decades. Additionally, we included the TRPM (Melastatin) subfamily and reviewed their role in cancer. Conclusively, these studies support that TRP channel-based therapies must be taken forward for clinical studies. Some channels, such as TRPC6, TRPM7 and TRPM8 were explored extensively in many cancer types which may be a potential target for cancer treatment. However, TRPM8 in lung cancer was reported for reverse association with cell proliferation, which needs to be reverified in lung cancer and other cancers. Besides, some TRPC channels are associated with store-operated calcium entry (SOCE) such as TRPC1, TRPC4 and TRPC6. Interestingly, the TRPC6 role was reported in breast cancer for modulation of Ca2 + through translocation of Orai1 and Orai3.","PeriodicalId":15286,"journal":{"name":"Journal of Cellular Biotechnology","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2022-07-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"42624413","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
M. Şamli, H. Samli, C. Gül, A. Ersoy, S. Ardıçlı, F. Balci
BACKGROUND: Semen analysis is a routine predictor of male fertility, and however, measurements of sperm morphology, motility, and concentration do not always evince genomic defects. OBJECTIVE: To investigate sperm parameters of renal transplant patients and to evaluate sperm DNA defects. METHODS: Seminal samples from 25 healthy controls and 56 transplantation patients were analyzed to evaluate DNA fragmentation by TUNEL. The differences in TUNEL-assay results and seminal parameters were compared between kidney transplant patients and controls. RESULTS: Among the azoospermic patients, 37.5% had fathered children before the disease. Three patients receiving sirolimus treatment had oligoasthenoteratozoospermia and infertility. In kidney transplant patients, DNA fragmentation was slightly higher than controls. Total motility (%) of the spermatozoa from the kidney transplant patients (42.2±21.9) was significantly lower (P < 0.05) than those of the control group (64.3±11.9). Moreover, control individuals had significantly higher (P < 0.05) normal morphology (23.2%) compared to the patient group (20.3%). Concerning sirolimus treatment, three patients had severe oligoasthenoteratozoospermia in their ejaculate, and however, DNA fragmentation rates were not significantly higher than those in the remaining individuals of the transplant group. CONCLUSIONS: The sperm DNA fragmentation rate in kidney transplant patients was slightly higher than in the control group (P = 0.09). However, the amount of spermatozoa DNA damage may lead to infertility in kidney transplant patients.
{"title":"TUNEL analysis of sperm DNA fragmentation in kidney transplant patients","authors":"M. Şamli, H. Samli, C. Gül, A. Ersoy, S. Ardıçlı, F. Balci","doi":"10.3233/jcb-220068","DOIUrl":"https://doi.org/10.3233/jcb-220068","url":null,"abstract":"BACKGROUND: Semen analysis is a routine predictor of male fertility, and however, measurements of sperm morphology, motility, and concentration do not always evince genomic defects. OBJECTIVE: To investigate sperm parameters of renal transplant patients and to evaluate sperm DNA defects. METHODS: Seminal samples from 25 healthy controls and 56 transplantation patients were analyzed to evaluate DNA fragmentation by TUNEL. The differences in TUNEL-assay results and seminal parameters were compared between kidney transplant patients and controls. RESULTS: Among the azoospermic patients, 37.5% had fathered children before the disease. Three patients receiving sirolimus treatment had oligoasthenoteratozoospermia and infertility. In kidney transplant patients, DNA fragmentation was slightly higher than controls. Total motility (%) of the spermatozoa from the kidney transplant patients (42.2±21.9) was significantly lower (P < 0.05) than those of the control group (64.3±11.9). Moreover, control individuals had significantly higher (P < 0.05) normal morphology (23.2%) compared to the patient group (20.3%). Concerning sirolimus treatment, three patients had severe oligoasthenoteratozoospermia in their ejaculate, and however, DNA fragmentation rates were not significantly higher than those in the remaining individuals of the transplant group. CONCLUSIONS: The sperm DNA fragmentation rate in kidney transplant patients was slightly higher than in the control group (P = 0.09). However, the amount of spermatozoa DNA damage may lead to infertility in kidney transplant patients.","PeriodicalId":15286,"journal":{"name":"Journal of Cellular Biotechnology","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2022-07-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"48069198","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Shivani Sachdeva, A. Dalvi, H. Saluja, Abhijeet Haridas, A. Mani
The currently prevalent COVID-19 infection, its line of treatment, resultant immunosuppression, and pre-existing comorbidities have made patients exposed to secondary infections including mucormycosis. Mucormycosis is a rare but in invasive fungal infection (IFI) due to several species of saprophytic fungi, occurring in patients with underlying co-morbidities which include diabetes mellitus, organ transplant, immunosuppressive corticosteroid therapy. The maxilla rarely undergoes necrosis due to its rich vascularity. Rare but not uncommon is the incidence of mucormycosis associated maxillary osteomyelitis occurring post COVID-19 infection. Fungal osteomyelitis is a life-threatening infection which may further spread from maxilla to the nose and paranasal sinuses within the orofacial region. It is an aggressive infection that needs to be addressed promptly to prevent fatal consequences.
{"title":"Coronavirus disease (COVID-19) Associated mucormycosis (CAM)","authors":"Shivani Sachdeva, A. Dalvi, H. Saluja, Abhijeet Haridas, A. Mani","doi":"10.3233/jcb-210046","DOIUrl":"https://doi.org/10.3233/jcb-210046","url":null,"abstract":"The currently prevalent COVID-19 infection, its line of treatment, resultant immunosuppression, and pre-existing comorbidities have made patients exposed to secondary infections including mucormycosis. Mucormycosis is a rare but in invasive fungal infection (IFI) due to several species of saprophytic fungi, occurring in patients with underlying co-morbidities which include diabetes mellitus, organ transplant, immunosuppressive corticosteroid therapy. The maxilla rarely undergoes necrosis due to its rich vascularity. Rare but not uncommon is the incidence of mucormycosis associated maxillary osteomyelitis occurring post COVID-19 infection. Fungal osteomyelitis is a life-threatening infection which may further spread from maxilla to the nose and paranasal sinuses within the orofacial region. It is an aggressive infection that needs to be addressed promptly to prevent fatal consequences.","PeriodicalId":15286,"journal":{"name":"Journal of Cellular Biotechnology","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2022-06-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"41325613","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Vida Mirzaie, Touba Eslaminejad, H. Babaei, S. Nematollahi-Mahani
BACKGROUND: Butyrylcholineesterase (BChE) is a therapeutic drug and its producing as a recombinant protein is an essential issue in biotechnology. One of the highlights in this regard is choosing the best host cells and plasmids. OBJECTIVES: The aim of this study is to evaluate the production of butyrylcholinesterase in Vero, HEK-293, and CHO cell lines using a dual promoter vector. MATERIAL AND METHODS: The dual-promoter construction (pBudCE dual BChE) was transfected into cell lines categorized in three experimental groups (pBudCE dual BChE, pCMV and negative control). BChE gene expression and enzyme activity was evaluated at different times. RESULTS: All three cell lines showed higher gene expression level in pBudCE dual BChE group. BChE enzyme activity level of this group in CHO cells decreased in sixth day and increased in ninth day. In HEK-293 cells it has a downward trend from sixth to ninth day and in Vero cells its level in the ninth day was the highest. CONCLUSION: The difference of pBudCE dual BChE and pCMV groups was more pronounced in the HEK-293 cell and the BChE gene expression level of this cells was higher than the others while, CHO cells showed higher level of BChE enzyme activity.
{"title":"Evaluation of the butyrylcholinesterase expression and activity in CHO, HEK-293 and vero cell lines transformed by dual promoter expression vector","authors":"Vida Mirzaie, Touba Eslaminejad, H. Babaei, S. Nematollahi-Mahani","doi":"10.3233/jcb-210042","DOIUrl":"https://doi.org/10.3233/jcb-210042","url":null,"abstract":"BACKGROUND: Butyrylcholineesterase (BChE) is a therapeutic drug and its producing as a recombinant protein is an essential issue in biotechnology. One of the highlights in this regard is choosing the best host cells and plasmids. OBJECTIVES: The aim of this study is to evaluate the production of butyrylcholinesterase in Vero, HEK-293, and CHO cell lines using a dual promoter vector. MATERIAL AND METHODS: The dual-promoter construction (pBudCE dual BChE) was transfected into cell lines categorized in three experimental groups (pBudCE dual BChE, pCMV and negative control). BChE gene expression and enzyme activity was evaluated at different times. RESULTS: All three cell lines showed higher gene expression level in pBudCE dual BChE group. BChE enzyme activity level of this group in CHO cells decreased in sixth day and increased in ninth day. In HEK-293 cells it has a downward trend from sixth to ninth day and in Vero cells its level in the ninth day was the highest. CONCLUSION: The difference of pBudCE dual BChE and pCMV groups was more pronounced in the HEK-293 cell and the BChE gene expression level of this cells was higher than the others while, CHO cells showed higher level of BChE enzyme activity.","PeriodicalId":15286,"journal":{"name":"Journal of Cellular Biotechnology","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2022-02-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"44400663","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
P. Kale, Amit Mani, Raju Anarthe, Rachita G Mustilwar
Tissue engineering aims to reconstruct the natural target tissue by a combination of three key elements stem/progenitor cells (that will create the new tissue), signaling molecules (that instruct the cells to form the desired tissue) scaffold/extracellular matrix (to hold the cells). Regeneration of the periodontal tissues following destructive episodes of various forms of periodontitis is a formidable challenge to periodontologists. Bone morphogenic proteins have been considered as the most potent growth factors that can promote the bone regeneration. This review will emphasize on the unique nature of the tissue engineered bone morphogenic proteins molecules regarding their structure, classification, signaling mechanism, etc. which will further help in understanding their role and potential advances necessary to facilitate the process of regeneration in the field of periodontics.
{"title":"Role of bone morphogenetic proteins in periodontal tissue engineering: Relatively unexplored horizon","authors":"P. Kale, Amit Mani, Raju Anarthe, Rachita G Mustilwar","doi":"10.3233/jcb-210051","DOIUrl":"https://doi.org/10.3233/jcb-210051","url":null,"abstract":"Tissue engineering aims to reconstruct the natural target tissue by a combination of three key elements stem/progenitor cells (that will create the new tissue), signaling molecules (that instruct the cells to form the desired tissue) scaffold/extracellular matrix (to hold the cells). Regeneration of the periodontal tissues following destructive episodes of various forms of periodontitis is a formidable challenge to periodontologists. Bone morphogenic proteins have been considered as the most potent growth factors that can promote the bone regeneration. This review will emphasize on the unique nature of the tissue engineered bone morphogenic proteins molecules regarding their structure, classification, signaling mechanism, etc. which will further help in understanding their role and potential advances necessary to facilitate the process of regeneration in the field of periodontics.","PeriodicalId":15286,"journal":{"name":"Journal of Cellular Biotechnology","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2021-12-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"42827380","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
BACKGROUND: Wound healing needs to occur after injury to prevent vision loss. Models of wound healing need to be optimized to assure treatments for corneal wounds can be developed in vitro prior to investigating with in vivo studies. OBJECTIVE: The purpose of this study was to establish the optimum media to use as a control solution in wound healing models. METHODS: Immortalized human corneal epithelial cells were cultured in different growth media using a scratch and exclusion zone model. The effect of normoxic and hypoxic conditions on tight junctional integrity and metabolic activity of cells grown in different growth medium were also investigated. RESULTS: Wound healing with DMEMF12 media was significantly faster than both Keratinocyte serum-free media (p < 0.05) and EpiLife (p < 0.05) after 10 hours recovery under normoxic or hypoxic conditions using the scratch model and 9 days after wounding using the exclusion zone technique (p < 0.05). Using the culture media DMEMF12, cells stained for abundant ZO-1, Cx43 and had a high metabolic activity indicating significant epithelial barrier formation, gap junction formation and high cell viability. CONCLUSIONS: DMEMF12 led to superior wound healing under hypoxic and normoxic conditions and in two different wound healing models.
{"title":"Establishment of optirmal culture media in corneal epithelial wound healing models","authors":"D. McCanna","doi":"10.3233/jcb-210039","DOIUrl":"https://doi.org/10.3233/jcb-210039","url":null,"abstract":"BACKGROUND: Wound healing needs to occur after injury to prevent vision loss. Models of wound healing need to be optimized to assure treatments for corneal wounds can be developed in vitro prior to investigating with in vivo studies. OBJECTIVE: The purpose of this study was to establish the optimum media to use as a control solution in wound healing models. METHODS: Immortalized human corneal epithelial cells were cultured in different growth media using a scratch and exclusion zone model. The effect of normoxic and hypoxic conditions on tight junctional integrity and metabolic activity of cells grown in different growth medium were also investigated. RESULTS: Wound healing with DMEMF12 media was significantly faster than both Keratinocyte serum-free media (p < 0.05) and EpiLife (p < 0.05) after 10 hours recovery under normoxic or hypoxic conditions using the scratch model and 9 days after wounding using the exclusion zone technique (p < 0.05). Using the culture media DMEMF12, cells stained for abundant ZO-1, Cx43 and had a high metabolic activity indicating significant epithelial barrier formation, gap junction formation and high cell viability. CONCLUSIONS: DMEMF12 led to superior wound healing under hypoxic and normoxic conditions and in two different wound healing models.","PeriodicalId":15286,"journal":{"name":"Journal of Cellular Biotechnology","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2021-12-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"42428255","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
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