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Reconstruction of maxillofacial defect still remains a challenge for oral and maxillofacial surgeons. Autograft, which has been the gold standard, has limitations and donor site morbidity. Tissue engineering combining cells, scaffold, and growth factors is expected to produce ideal bone without morbidity, with Human umbilical cord mesenchymal stem cells (HUCMSC) and bovine scaffold are promising topics. HUCMSC therapy is influenced by biomolecules including cytokines, growth factors, and microRNAs, known as secretome. This study aims to analyze the osteoinduction potential of the HUCMSC secretome in the FDBBX scaffold through the expression of Runt Related Transcription Factor-2 (RUNX2) and Osteocalcin (OCN) markers. Objectives to analyze the increased expression of RUNX2 and OCN of MC3T3-E1 preosteoblast cells that were seeded into FDBBX scaffold after application of the HUCMSC secretome compared to scaffold without secretome. The study was an in vitro laboratory experiment. The first group was FDBBX scaffold after HUCMSC secretome application and the second group was FDBBX without secretome. The analysis of the RUNX2 and OCN expression of MC3T3-E1 pre-osteoblast cells that attached to the scaffold was conducted on days 7, 14 and 21 using RT-PCR method. Independent T test was conducted as a Statistical test to analyze the differences between groups, and One way ANOVA test was conducted to analyze the difference between time. The difference analysis was considered significant when p<0,05. The Independent T test result showed that the expression of RUNX2 and OCN in FDBBX scaffold after HUCMSC secretome application was significantly higher than scaffold without secretome (P=0,039; P=0,025). The expression of RUNX2 and OCN of MC3T3-E1 pre-osteoblast cells in the FDBBX scaffold after HUCMSC secretome application was higher than the scaffold without secretome application. [ABSTRACT FROM AUTHOR]

A potent therapy for the infectious coronavirus disease COVID-19 is urgently required with, at the time of writing, research in this area still ongoing. This study aims to evaluate the in vitro anti-viral activities of combinations of certain commercially available drugs that have recently formed part of COVID-19 therapy. Dual combinatory drugs, namely; Lopinavir-Ritonavir (LOPIRITO)-Clarithromycin (CLA), LOPIRITO-Azithromycin (AZI), LOPIRITO-Doxycycline (DOXY), Hydroxychloroquine (HCQ)-AZI, HCQ-DOXY, Favipiravir (FAVI)-AZI, HCQ-FAVI, and HCQ-LOPIRITO, were prepared. These drugs were mixed at specific ratios and evaluated for their safe use based on the cytotoxicity concentration (CC50) values of human umbilical cord mesenchymal stem cells. The anti-viral efficacy of these combinations in relation to Vero cells infected with SARS-CoV-2 virus isolated from a patient in Universitas Airlangga hospital, Surabaya, Indonesia and evaluated for IC50 24, 48, and 72 hours after viral inoculation was subsequently determined. Observation of the viral load in qRT-PCR was undertaken, the results of which indicated the absence of high levels of cytotoxicity in any samples and that dual combinatory drugs produced lower cytotoxicity than single drugs. In addition, these combinations demonstrated considerable effectiveness in reducing the copy number of the virus at 48 and 72 hours, while even at 24 hours, post-drug incubation resulted in low IC50 values. Most combination drugs reduced pro-inflammatory markers, i.e. IL-6 and TNF-α, while increasing the anti-inflammatory response of IL-10. According to these results, the descending order of effective dual combinatory drugs is one of LOPIRITO-AZI>LOPIRITO-DOXY>HCQ-AZI>HCQ-FAVI>LOPIRITO-CLA>HCQ-DOX. It can be suggested that dual combinatory drugs, e.g. LOPIRITO-AZI, can potentially be used in the treatment of COVID-19 infectious diseases. [ABSTRACT FROM AUTHOR]

Introduction: The preconditioning of HAMSCs using Cobalt (II) Chloride (CoCl2) as a hypoxia mimicking agent (HMA) may provide the adaptive state of HAMSCs. Objective: To examine the effect of CoCl2 in the viability percentage and the induced Hypoxia Inducible Factor-1α (HIF-1α) expression of HAMSCs in vitro. Methods: HAMSCs were thawed, cultured, and then sub-cultured from the laboratory cell stock. The Immunocytochemistry (ICC) methods with Fluorescein isothiocyanate (FITC) antibody labelling kit was used to validate the HAMSCs employing its Cluster of Differentiation (CD) expression such as CD73(+), CD105(+) and CD45(-). HAMSCs were sub-cultured and then co-cultivated with or without CoCl2 in the different concentration (200 (U+03BC)mol, 150 (U+03BC)mol, 100 (U+03BC)mol, 75 (U+03BC)mol, 50 (U+03BC)mol, 25 (U+03BC)mol) for 24 hours. The cytotoxicity test was conducted by utilizing tetrazolium dye MTT 3-(4,5- dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide. The formazan crystal was then observed, and the cell viability percentage was calculated. In examining CoCl2 induced hypoxia in HAMSCs, the expression of HIF-1α applied ICC-FITC in vitro. Analysis of Variance (ANOVA) was performed to analyze the obtained data statistically (p<0.05). Results: HAMSCs express CD73 and C105 positively, while HAMSCs expressed CD45 negatively. There is no significant different in cell viability percentage of HAMCs between groups with different concentrations of CoCl2 (p=0.99; p>0.05). CoCl2 successfully induces the hypoxia confirmed by HIF-1α positive expression of HAMSCs after 24 hours incubation in vitro. Conclusion: CoCl2 does not affect the viability percentage of HAMSCs and it can be used as a biocompatible HMA in HAMSCs in vitro. [ABSTRACT FROM AUTHOR]

Previous research focuses on in vitro study of stem cell metabolites derived from placenta for wound healing. This study, however, is an advanced stage which focuses on testing the efficiency and efficacy of stem cell metabolites in rats (Rattus novergicus). The tests carried out examined the blood levels with ELISA instruments and integument histology by observing the activity of polymorph nuclear and monocyte cells in the control and treatment groups. In the control group, the rats were injured in the anterior and posterior back skin with a 1×1 cm incision wound, (only antibiotics), while the treatment group uses antibiotics and 4 mL injections of stem cell metabolites. Each group was repeated three times with the samples observed for blood levels using ELISA Interleukin-4, Interleukin-10 and Tumor Necrosis Factor-α, with integument histology at pre-injection, in days 1, 3 and 6. These were used to compare the development of inflammatory cells, polymorphonuclear and monocytes between the control and treatment groups. Stem cell metabolites are significantly effective and efficient with the ability to inhibit the inflammatory process in tissues in terms of examining the blood levels of rats using ELISA Interleukin-4, Interleukin-10 and Tumor Necrosis Factor-α. Interleukin-4 and Interleukin-10 (anti-inflammatory) tend to significantly increase the treatment group, while Tumor Necrosis Factor-α (pro-inflammation) increases the control group. Histology results showed a decrease in the activity of polymorphonuclear and monocytes inflammatory cells in the treatment group compared to the control, which indicated that the stem cell metabolites were able to significantly inhibit the inflammatory process. It is concluded that stem cell metabolites derived from placenta are effective and efficient for wound healing in rats. Clinical study is needed for further research for it to be used on humans. [ABSTRACT FROM AUTHOR]

Mesenchymal stem cells (MSCs) are undifferentiated cells with potential for therapeutic use in degenerative disease, trauma, or injury. In this study, wounds on the dorsal skin of mice resulting from an iron scaffold were injected with 105 activated MSCs. Crude secretions obtained from maintenance stem cell medium was labeled using green fluorescent protein following an intravenous lateral tail administration. Bioluminescent imaging to determine the MSC migration into the cutaneous burn injury site was performed after 0, 24 and 48 hours. The results obtained suggest that active MSCs accelerate the burn healing process. [ABSTRACT FROM AUTHOR]

Rabbits (Oryctolagus cuniculus) are widely used as experimental models for both human and veterinary medicine because of their ease to work with and relatively economic maintenance. Rabbits are useful for multiple applications in biotechnology and translational medicine from basic research to preclinical studies. Skin has been considered a potential source of adult stem cells. Skin-Derived Mesenchymal Stem Cells (SMSCs) are an abundant pluripotent, multipotent cell source and capable of undergoing in vitro osteogenic differentiation. The differentiation of SMSCs in vitro is induced by the presence of dexamethasone, ascorbic acid and β-glycerol phosphate. The characterization of Skin-Derived Mesenchymal Stem Cells (SMSCs) showed the expression of CD105, CD73, CD90 for positive markers, as well as CD45 for negative markers using immunofluorescence methods for mesenchymal stem cells (MSCs). The present work first determined the osteogenic differentiation from SMSCs for 21 days, then confirmed the characterized expression of osteocalcin, runt-related transcription factor 2 (Runx2), SRY-related HMG-box (Sox9) with immunofluorescence and analyzed by alizarin red staining for osteogenic differentiation. [ABSTRACT FROM AUTHOR]

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Purpose: To analyze the expression of bone sialoprotein - I (BSP - I) after the addition of platelet rich fibrin (PRF) in gingival somatic cell (GSC) culture medium during osteogenic differentiation in vitro. Methods: GSCs were extracted from healthy, 1-month-old, male Wistar rats (Rattus Novergicus), weighing 250 - 300 g, and which had been randomly selected (n=4). These cells were cultured for 14 days and passaged every 4 days. Five subcultures of GSCs were cultured in three plates (M24) (N = 54; n = 6) for 7, 14 and 21 days in three preconditioned culture media (group I: plain culture media; group II: preconditioned osteogenic culture media, and group III: preconditioned osteogenic culture media with platelet rich fibrin). The expression of BSP-I was immunocytochemically (ICC) examined with monoclonal antibodies. Homogeneity and normality tests (p > 0.05) were then performed followed by an analysis of variance (ANOVA, p < 0.05). Results: The highest expression of BSP-I was found in group III (Day 21, 13.00 ± 2.000), while the lowest expression of BSP-I was found in group I (Day 7, 7.33 ± 1.155). There were significant differences between the groups (p = 0.000, p < 0.05). Conclusion: PRF stimulates and significantly enhances the expression of BSP-I in GSC culture during osteogenic differentiation. Thus, PRF can be used to accelerate regeneration of alveolar bone defects. [ABSTRACT FROM AUTHOR]

Background: At the present time, COVID-19 vaccines are at the testing stage, and an effective treatment for COVID-19 incorporating appropriate safety measures remains the most significant obstacle to be overcome. A strategic countermeasure is, therefore, urgently required., Aim: This study aims to evaluate the efficacy and safety of a combination of lopinavir/ritonavir-azithromycin, lopinavir/ritonavir-doxycycline, and azithromycin-hydroxychloroquine used to treat patients with mild to moderate COVID-19 infections. Setting and Design . This study was conducted at four different clinical study sites in Indonesia. The subjects gave informed consent for their participation and were confirmed as being COVID-19-positive by means of an RT-PCR test. The present study constituted a randomized, double-blind, and multicenter clinical study of patients diagnosed with mild to moderate COVID-19 infection., Materials and Methods: Six treatment groups participated in this study: a Control group administered with a 500 mg dose of azithromycin; Group A which received a 200/50 mg dose of lopinavir/ritonavir and 500 mg of azithromycin; Group B treated with a 200/50 mg dose of lopinavir/ritonavir and 200 mg of doxycycline; Group C administered with 200 mg of hydroxychloroquine and 500 mg of azithromycin; Group D which received a 400/100 mg dose of lopinavir/ritonavir and 500 mg of azithromycin; and Group E treated with a 400/100 mg dose of lopinavir/ritonavir and 200 mg of doxycycline., Results: 754 subjects participated in this study: 694 patients (92.4%) who presented mild symptoms and 57 patients (7.6%) classified as suffering from a moderate case of COVID-19. On the third day after treatment, 91.7%-99.2% of the subjects in Groups A-E were confirmed negative by a PCR swab test compared to 26.9% in the Control group. Observation of all groups which experienced a significant decrease in virus load between day 1 and day 7 was undertaken. Other markers, such as CRP and IL-6, were significantly lower in all treatment groups ( p < 0.05 and p < 0.0001) than in the Control group. Furthermore, IL-10 and TNF- α levels were significantly elevated in all treatment groups ( p < 0.0001). The administration of azithromycin to the Control group increased CRP and IL-6 levels, while reduced IL-10 and TNF- α on day 7 ( p < 0.0001) compared with day 1. Decreases in ALT and AST levels were observed in all groups ( p < 0.0001). There was an increase in creatinine in the serum level of the Control, C, D, and E groups ( p < 0.05), whereas the BUN level was elevated in all groups ( p < 0.0001)., Conclusions: The study findings suggest that the administration of lopinavir/ritonavir-doxycycline, lopinavir/ritonavir-azithromycin, and azithromycin-hydroxychloroquine as a dual drug combination produced a significantly rapid PCR conversion rate to negative in three-day treatment of mild to moderate COVID-19 cases. Further studies should involve observation of older patients with severe clinical symptoms in order to collate significant amounts of demographic data., Competing Interests: The authors declare no conflicts of interest regarding this work., (Copyright © 2021 Purwati et al.)