Cytokine profiles of mild-to-moderate SARS-CoV-2 infected and recovered pre-vaccinated individuals residing in Indonesia.

Background: Accumulating evidence suggests the involvement of cytokine-mediated inflammation, in clinical severity and death related to SARS-CoV-2 infection, especially among pre-vaccinated individuals. An increased risk of death was also described among SARS-CoV-2 recovered individuals, which might be correlated with prolonged inflammatory responses. Despite being among the countries with the highest cumulative deaths due to COVID-19, evidence regarding cytokine profiles among SARS-CoV-2 infected and recovered pre-vaccinated individuals in Indonesia is scarce. Thus, this study aimed to describe the cytokines profiles of pre-vaccinated individuals residing in Indonesia, with mild-to-moderate SARS-CoV-2 infection and those who recovered. Methods: Sixty-one sera from 24 hospitalized patients with mild-to-moderate SARS-CoV-2 infection, 24 individuals recovered from asymptomatic-to-moderate SARS-CoV-2 infection, and 13 healthy controls unexposed to SARS-CoV-2 were used in this study. Quantification of serum cytokine levels, including IL-6, IL-8, IP-10, TNF-α, CCL-2, CCL-3, CCL-4, and CXCL-13, was performed using a Luminex multi-analyte-profiling (xMAP)-based assay. Results: The levels of IL-8 along with CCL-2 and CCL-4, were significantly higher (p ≤ 0.01) in hospitalized patients with mild-to-moderate SARS-CoV-2 infection and recovered individuals compared to healthy controls. However, no significant difference was observed in these cytokine levels between infected and recovered individuals. On the other hand, there were no significant differences in several other cytokine levels, including IL-6, IL-10, TNF-α, CCL-3, and CXCL-13, among all groups. Conclusion: IL-8, CCL-2, and CCL-4 were significantly elevated in pre-vaccinated Indonesian individuals with mild-to-moderate SARS-CoV-2 infection and those who recovered. The cytokine profiles described in this study might indicate inflammatory responses not only among SARS-CoV-2 infected, but also recovered individuals.

In-Vitro Inhibitory Efficacy of 3 Types of Probiotics on the Growth of Aggregatibacter actinomycetemcomitans Bacteria.

BACKGROUND: Amongst the specific plaque pathogen Aggregatibacter actinomycetemcomitans (Aa) ATCC 43718 serotype b is one of the highly virulent bacteria that causes periodontitis. Probiotic therapy is a treatment in which the lactic acid bacteria in are utilized to impede the colonization and growth of the pathogenic bacteria to prevent the further formation of dental plaque. OBJECTIVE: The present research aimed to evaluate inhibiting effect of purified bacteria from various commercially available yogurt product containing bacteria named (Lactobacillus casei strain Shirota; Lactobacillus bulgaricus and Streptococcus thermophilus; Lactobacillus reuteri Prodentis) on the growth of Aa. METHODS: The research made use of the diffusion method by fixing Aa on BHIB (brain heart infusion broth) medium, incubated at 37 °C and 24 hours later planted on MHA (Mueller-Hinton agar) media. Aa were divided into four subgroups each with a paper disk; group 1 consists of untreated bacteria (i.e., control group), group 2 with purified bacteria from Yakult 0.5 µL, group 3 with purified bacteria from Cimory Yogurt Drink 0.5 µL and group 4 with purified bacteria from BioGaia Prodentis 0.5 µL. All commercially available yogurt were treated to get the purified probiotic. Additionally, it was incubated for 24 hours at 37 °C and later the inhibition zone diameter was observed. RESULTS: In the research, it was found that the average impeding ability, so-called inhibition zone, in group 1 indicated 0 mm, group 2 indicated 12.70 mm, group 3 indicated 16.60 mm and group 4 indicated 19.60 mm. The statistical test outcomes showed a significance of 0.000 (p < 0.05). CONCLUSIONS: The purified bacteria from three probiotics indeed inhibit the growth of the Aa bacteria and a substantial difference in the diameter of the inhibition zone were found among the three probiotics.

RUNX2 and ALP expression in osteoblast cells exposed by PMMA-HAp combination: An in vitro study.

Objective: To observe the expression of Runt-Related Transcription Factors 2 (RUNX2) and Alkaline Phosphatase (ALP) markers in osteoblast cell cultures exposed to Polymethylmethacrylate (PMMA) combined with hydroxyapatite (HAp) material to improve osteointegration of bone implants. Methods: Sample of PMMA and HAp materials with a mixture of PMMA with HAp made from limestone as natural source which processed through Balai Besar Keramik (HApBBK) in the first group and a mixture of PMMA with HAp made from bovine bone which processed through Good Manufacturing Practice (HApGMP) in the second group. Twenty-four fetal rat calvarie osteoblast cell cultures were randomly divided into 6 groups: 7- and 14-day control group, 7 and 14 days PMMA-HApGMP group, 7 and 14 days PMMA-HApBBK group. The expression of RUNX2 and ALP was seen by immunocytochemical examination. Result: The one-way ANOVA with a significance value of 0.000 (p < 0.05). There was an increase in RUNX2 and ALP expressions on both PMMA-HApBBK and PMMA-HApGMP groups on days 7 and 14 in osteoblast cell cultures. Conclusion: The PMMA-HApBBK and PMMA-HApGMP showed an increase in the RUNX2 and ALP expression in osteoblast cell cultures which indicates a potential increase of osseointegration of bone implants.

Expression of VEGF and BMP-2 in Osteoblast cells exposed to a combination of polymethylmethacrylate (PMMA) and hydroxyapatite (HAp).

Objectives: Polymethylmethacrylate (PMMA) has been widely used, but it has several fallback properties in its interaction with bone tissue, so the addition of hydroxyapatite (HAp) material aims to improve the biocompatibility, regeneration process, and osteointegration of bone implants. The HAp material can be sourced from bovine bone and processed through Good Manufacturing Practice from Tissue Bank (HApGMP), and from limestone (CaCO3) processed by Balai Besar Keramik (HApBBK).This study was to observe the expression of vascular endothelial growth factor (VEGF) and Bone morphogenetic protein-2 (BMP2) in cultured osteoblasts exposed to PMMA-HApGMP and PMMA-HApBBK as implant candidate materials. Methods: Sample of PMMA and HAp materials with a mixture of PMMA and HApBBK in the first group and a mixture of PMMA and HApGMP in the second group. Twenty-four fetal rat calvarie osteoblast cell cultures were randomly divided into 6 groups: 7- and 14-day control group, 7 and 14 days PMMA-HApGMP group, 7 and 14 days PMMA-HApBBK group. The expression of VEGF and BMP-2 was seen by immunocytochemical examination. Results: The one-way ANOVA with a significance value of 0.000 (p < 0.05). BMP-2 and VEGF expression was increased in the 7- and 14-days groups after exposure to PMMA-HApGMP and PMMA-HApBBK. Conclusion: The application of PMMA-HApGMP and PMMA-HApBBK showed an increase in the expression of VEGF and BMP-2 in osteoblast cell cultures which indicates a potential increase in the accelerated angiogenesis and osteogenesis in the bone regeneration process of bone implants.

Concanavalin A Enhanced Proliferation and Osteogenic Differentiation of Dental Pulp Stem Cells.

OBJECTIVE: Dental pulp stem cells (DPSCs) can be used as a component in the formation of regenerative dentine during direct pulp capping therapy. Concanavalin A (ConA) is a type of lectin with a molecular weight of 26 kDa derived from the Canavalia ensiformis plant. Lectins possess strong proliferation and differentiation abilities in various animal cells including lymphocytes, osteoblasts, and chondrocytes. The aim of study was to determine the effect of ConA on the proliferation and osteogenic differentiation of DPSCs in vitro. MATERIALS AND METHODS: In this in vitro study, DPSCs were isolated from third molars before ConA induction was performed at concentrations of 5 and 10 µg/mL. The proliferation assay was determined by 3-(4,5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT) assay. Osteogenic differentiation was determined by means of mineralization. STATISTICAL ANALYSIS: Data were analyzed using analysis of variance and a Student's t-test. The p-value was set at 0.05. RESULTS: The addition of 5 and 10 µg/mL of ConA to DPSCs can significantly increase the proliferation and osteogenic differentiation of DPSCs (p ≤0.05). CONCLUSION: ConA can increase the proliferation and osteogenic differentiation of DPSCs.