Journal of Cellular Biotechnology - Volume 8, issue 2

Authors: Nguyen, Thao Thi Thanh | Ta, Giang Huong | Le, Le Thi Thu | Vo, Thao Thanh Nguyen | Pham, Tram Thi Kim | Nguyen, Quan Dang

Article Type: Research Article

Abstract: BACKGROUND: TNF-α (Tumor necrosis factor-alpha) plays a vital role in the human immune system. However, abnormal activity and overexpression of TNF-α are some of the causes of allergic and autoimmune diseases. Inhibiting the activity of this molecule is one of the novel pathologies for these diseases. The development of this recombinant protein is expected to reduce the financial burden of treating inflammatory rheumatic diseases. OBJECTIVE: The study’s objective was to generate and evaluate the biological activity of TNFR-Fc, construct of fusions an extracellular part TNF-α receptor (p75) and an Fc fragment of human immunoglobulin G1, expressed from the …CHO-DG44 cell system. METHODS: The recombinant TNFR- Fc plasmid was constructed and identified by PCR, restriction enzyme digestion, and sequencing. A stable cell line for expression of TNFR-Fc was selected by limiting dilution cloning. Structural characterization, the binding affinity of TNFR-Fc to TNF-α, the neutralizing the cytotoxic activity- induced TNF-α, and the TNF-α-induced apoptosis suppression of TNFR- Fc were analyzed by SDS/PAGE Western blotting, ELISA, WST assay, Immunofluorescence, and flow cytometry. RESULTS: Preliminary analysis of the structural characteristics showed that TNFR-Fc is a low- glycosylated protein and perhaps in dimeric form. Furthermore, the recombinant TNFR-Fc can interact with its ligand TNF-α with a dissociation constant Kd 0.25±0.03μM equivalent to that of the original drug, Etanercept. We also demonstrated that TNFR-Fc expressed from CHO-DG44 was able to neutralize TNF-α- induced cytotoxic activity and inhibited p53-related apoptosis in vitro , similar to Etanercept. CONCLUSIONS: These data collectively suggested that TNFR-Fc potently blocks TNF-α, which could be a novel therapeutic strategy for cytokine-driven diseases. Show more

Keywords: TNF-α, TNFR-Fc, etanercept, CHO-DG44

DOI: 10.3233/JCB-210048

Citation: Journal of Cellular Biotechnology, vol. 8, no. 2, pp. 51-66, 2022

Authors: Rao, Subramaniam M. | Naresh Kumar, C. | Kokatnur, Vijayalaxmi | Rao, Anupama | Singh, Varsha

Article Type: Research Article

Abstract: Reactive oxygen species (ROS) are free radical molecules obtained from biotransformation of molecular oxygen. Oxidative stress is referred as oxidative damage produced by free radicals, and has been associated with many diseases including periodontitis. Periodontitis is a chronic inflammatory disease resulting in loss of attachment and bone loss. There is over production of ROS in periodontitis, imbalance between free radicals and antioxidants locally result in destruction of periodontal tissues. Flavonoids are naturally occurring compounds known for various actions, such as, anti-inflammatory, antioxidant properties. The objective of this review is to provide an outline on the biological actions of flavonoids and …their beneficial effects on periodontal tissues. Show more

Keywords: Anti-oxidants, flavonoids, periodontitis, ROS

DOI: 10.3233/JCB-210062

Citation: Journal of Cellular Biotechnology, vol. 8, no. 2, pp. 67-74, 2022

Authors: Verma, Priyanka | Rani, Rachana | Rao, Priya | Singh, Anuj Kumar

Article Type: Research Article

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. Show more

Keywords: Ion channel, TRP channels, Cancer, Migration, acidic extracellular pH

DOI: 10.3233/JCB-220066

Citation: Journal of Cellular Biotechnology, vol. 8, no. 2, pp. 75-84, 2022

Authors: Samli, Murat | Samli, Hale | Gul, Cuma Bulent | Ersoy, Alparslan | Ardicli, Sena | Balci, Faruk

Article Type: Research Article

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. Show more

Keywords: Immunosuppressive treatment, kidney transplantation, spermatogenesis, sperm DNA damage, TUNEL analysis

DOI: 10.3233/JCB-220068

Citation: Journal of Cellular Biotechnology, vol. 8, no. 2, pp. 85-93, 2022

Authors: Sachdeva, Shivani | Mani, Amit | Vora, Hiral | Saluja, Harish | Manka, Nishant | Dehane, Vipin

Article Type: Research Article

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. Show more

Keywords: Tissue engineering, scaffolds, biomaterial, bioprinting, regeneration

DOI: 10.3233/JCB-220070

Citation: Journal of Cellular Biotechnology, vol. 8, no. 2, pp. 95-103, 2022

Authors: Pendyala, Gowri | Joshi, Saurabh | Mani, Ameet | Dhole, Sudhir | Kale, Preeti

Article Type: Research Article

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. Show more

Keywords: Nonsurgical periodontal therapy, scaling and root planing, ozone oil, chlorhexidine gel, toxicity, antimicrobial, inflammatory mediator

DOI: 10.3233/JCB-220073

Citation: Journal of Cellular Biotechnology, vol. 8, no. 2, pp. 105-112, 2022