Analysis of Saliva Composition: Parathyroid Hormone-Related Protein, Total Protein, and Secretory Immunoglobulin A (sIgA) in Rattus norvegicus with Stunted Growth.

OBJECTIVE: This study aimed to determine total protein, secretory immunoglobulin A (sIgA) and parathyroid hormone-related protein (PTHrP) levels in the saliva of rats with stunted growth. MATERIALS AND METHODS: Experimental laboratory research with a pre-and posttest control group design was conducted. Seventeen albino rats (Rattus norvegicus) were divided into the control group (eight rats) and the treatment group (nine rats). Rats in the treatment group were exposed to aflatoxin B1 5µg/kg orally for 5 weeks. Anthropometry data (body length, body weight) and saliva of R. norvegicus were collected. The levels of PTHrP and sIgA in the saliva were measured using an enzyme-linked immunosorbent assay kit for rats and the Bradford test for total protein and analyzed using SPSS 25.0. RESULTS: Aflatoxin caused stunted growth in rats in the treatment group. There was a significant difference in body length, salivary flow, PTHrP, sIgA, and total protein in the treatment group compared with the control group. The average rat's body length change in the control group was 6.4 ± 1.1mm/5 weeks, while in the treatment group, the change was 3.7 ± 0.9 mm /5 weeks. There was no significant weight gain in the treatment group compared with the normal group. The average values of PTHrP, sIgA, and total protein in the control group were x̄0.9, x̄18, and x̄0.7 m./L, respectively, while in the treatment group, they measured x̄0.4, x̄10.7, and x̄0.5 mg/L, respectively. CONCLUSION: This study showed that salivary flow, PTHrP, sIgA, and total protein levels in the saliva were significantly lower in stunted rats compared with normal rats.

Pigmented Fungiform Papillae (PFP) of the Tongue: A Systematic Review of Current Aetiopathogenesis and Pathophysiology.

The pigmentation of the fungiform papillae of the tongue is a rare idiopathic condition in which only the fungiform papillae appear hyperpigmented. In the absence of any reviews on the subject, we conducted a systematic review of the aetiopathogenesis and pathophysiology of pigmented fungiform papillae (PFP) of the tongue, including its demographic and histopathological features, trying to outline a possible aetiology. The preferred reporting items for systematic reviews and meta-analyses (PRISMA) was performed using PubMed, Scopus, EMBASE databases and manual searches, for publications between January 1974 and July 2022. Inclusion criteria were case reports defining patients' characteristics, their general medical and dental conditions, histopathological and/or immunohistochemical findings, all with a final definitive diagnosis of PFP. Overall, 51 studies comprising 69 cases of PFP which included histopathological descriptions were reviewed. Prominent features consisted of hyperpigmentation of melanocytes, melanophages, chromatophores, and a lymphocytic infiltrate in the subepidermal area of the fungiform papillae. On special staining, PFP contained melanin, not iron or hemosiderin. On immunohistochemistry, immune-reactive CD3+ T lymphocytes, S-100 and Sox10, but non-immune-reactive melan-A intraepithelial melanocytes were noted in some studies. The presence of hyperpigmented melanocytes and melanophages, with non-immune-reactive melan-A, suggests that PFP are a benign and physiological form of pigmentation. The inflammatory infiltrates described in some papillary lesions could possibly be due to traumatic events during mastication. Nevertheless, the true reasons for the hyperpigmentation of the fungiform papillae are as of yet elusive, and remain to be determined.

Validation of RNA Aptamer Probes to Image Candida albicans in Paraffin-Embedded Sections of Wistar Rat Tongue.

OBJECTIVE: This study aimed to validate the use of Ca-apt-1, an RNA aptamer, that we generated previously as a probe for immunostaining of Candida albicans in rat tongue paraffin-fixed tissue sections MATERIAL AND METHODS: The performance of Ca-apt-1 as a detector molecule was compared with that of anti-C. albicans polyclonal antibody (PcAb), which was used as a positive control. Immunostaining images were visualized by light microscopy and were analyzed by using ImageJ software. RESULTS: Microscopic results demonstrated that Ca-apt-1 specifically recognized and immunostained C. albicans cells of rat tongue candidiasis, with a specificity comparable to that of PcAb. ImageJ analysis showed that the area (pixels) detected by Ca-apt-1 was wider than that detected by the antibody. This indicates that the binding affinity of Ca-apt-1 toward C. albicans was better than that of PcAb on paraffin-embedded tissues. CONCLUSION: This study demonstrates that Ca-apt-1 can be used as a probe for immunostaining of fixed tissue sections for oral candidiasis diagnosis.

The Effect of Lipoteichoic Acid from Lactobacillus plantarum on Dental Pulp Inflammation.

OBJECTIVE: Lactobacillus plantarum, a bacterium located in deep caries, has a virulence factor in the form of lipoteichoic acid (LTA), which is found in the bacterial cell wall. LTA is able to trigger a neutrophils response in the dental pulp inflammation process within the first 6 to 24 hours. The quantity of bacteria is one factor influencing the increase in number of neutrophils in addition to the quality of the bacteria. This study seeks to analyze the effect of lipoteichoic acid of Lactobacillus plantarum (LTA-Lp) in the dental pulp inflammation by observing the number of neutrophil cells in a histopathological view. MATERIALS AND METHODS: The LTA was isolated from L. plantarum. The left upper molar of Rattus novergicus was mechanically perforated under anesthesia to induce dental pulp inflammation. The perforated tooth was then induced by 10 and 15 µg/mL of LTA-Lp and then restored by a temporary filling. The perforated tooth in the control group was only restored by a temporary filling. After 24, 48, and 72 hours, the tooth was extracted and then stained with hematoxylins and eosin to observe the neutrophils in the dental pulp via a light microscope. RESULT: The number of neutrophils in the dental pulp after induction by 15 µg/mL of LTA-Lp is higher than 10 µg/mL of LTA-Lp and both controls. There were significant differences in the number of neutrophils in the dental pulp, in each group on 24, 48, and 72 hours after LTA-Lp inducing (p < 0.05). CONCLUSION: The LTA-Lp dose of of 10 and 15 µg/mL affected the dental pulp inflammation by affecting the number of neutrophils.

The Effect of Anadara granosa Shell's-Stichopus hermanni Scaffold on CD44 and IL-10 Expression to Decrease Osteoclasts in Socket Healing.

OBJECTIVES: This article aimed to investigate the effect of Anadara granosa (AG) shell's-Stichopus hermanni scaffold on cluster of differentiation (CD)44 and interleukin-10 (IL-10) expression to decrease osteoclasts in socket healing. MATERIALS AND METHODS: Thirty male Wistar rats were divided into five groups. The lower left incisor was extracted, then given a placebo for group control (K), the treatment group was administered with scaffold from AG shells, and a treatment group with scaffold from blood cockle shell-S. hermanni with the concentration of 0.4, 0.8, and 1.6% (AGSH0.4; AGSH0.8; AGSH1.6). We made a bone graft from a combination of AGSH extract using the freeze-dried method. The socket was sutured by silk braid immediately. Third and Seventh days postextraction, animals are killed. CD44 and IL-10 expression were examined with immunohistochemistry, as well as osteoclast was examined with hematoxylin-eosin. STATISTICAL ANALYSIS: The data were analyzed using a one-way analysis of variance (for CD44 and osteoclast) and Kruskal-Wallis' test (for IL-10) followed by a post hoc test in which the result of p < 0.05. RESULTS: Scaffold from a combination of AGSH increased CD44 expression significantly, which enhanced IL-10 expression thereby decreased the number of osteoclasts in socket healing on days 3 and 7. CONCLUSION: Scaffold of AG shell-S. hermanni with a concentration of 0.8% was effective to enhance CD44 and IL-10 expression to decrease osteoclast in socket healing after tooth extraction.

Detection of human immunodeficiency virus type 1 transmitted drug resistance among treatment-naive individuals residing in Jakarta, Indonesia.

The presence of transmitted drug resistance (TDR) in human immunodeficiency virus type 1 (HIV-1) infected individuals naive to antiretroviral therapy, may affect the effectiveness of treatment. Jakarta, the capital city of Indonesia, recorded the highest number of cumulative HIV infection cases in the country. This study aimed to identify on the appearance of TDR, as well as to identify HIV-1 subtypes circulating among treatment-naive individuals in Jakarta. Whole blood samples collected from 43 HIV-1 infected, treatment-naive individuals. Viral subtyping and drug resistance testing were performed on HIV-1 pol genes amplified using nested polymerase chain reaction. CRF01_AE was detected most frequently in Jakarta (73.08%). Drug resistance-related major mutation was not detected in protease fragments of pol gene, but two major mutations, K103N (6.67%) and Y181C (6.67%), were detected in reverse transcriptase fragments of pol gene. Our results suggest that TDR was emerged in Jakarta at a certain extent, thus further surveillance study to monitor the TDR prevalence and circulating HIV-1 subtypes in this region is considered to be necessary.

Combinations of propolis and Ca(OH)2 in dental pulp capping treatment for the stimulation of reparative dentin formation in a rat model.

Background: Caries in the dental pulp result in inflammation and damage to the pulp tissue. During inflammation of the pulp, various inflammatory mediators and growth factors are released, including IL-8, IL-10, TLR-2, VEGF and TGF-ß through the NF-kB pathway. In the present study, therapy for pulpal caries was performed through pulp capping by giving a combination of propolis and calcium hydroxide (Ca(OH)2). This treatment was expected to stimulate the formation of reparative dentin as an anti-inflammatory material to prevent pulp tissue damage. Methods: 28 Wistar rats were divided into four groups and treated with Ca(OH)2 with or without the addition of propolis for either 7 or 14 days. Immunohistochemical examination was used to determine the expression of IL-8, IL-10, TLR-2, VEGF, TGF-ß in the four treatment groups. Results: The group treated with a combination of propolis and Ca(OH)2 for 7 days showed that the expression of IL-10, IL-8, TLR-2, VEGF, TGF-ß increased significantly compared to the treatment group treated with only Ca(OH)2. The expression of IL-10, TLR-2, TGF-ß, VEGF increased in the treatment group treated with propolis and Ca(OH)2 for 14 days, while the expression of IL-8 in the decreased significantly. Conclusions: Administration of a combination of propolis and Ca(OH)2 has efficacy in the pulp capping treatment process because it has anti-bacterial and immunomodulatory properties. The results show that it is able to stimulate the process of pulp tissue repair through increased expression of IL-10, TGF-ß, VEGF, TLR -2 and decreased expression of IL-8.

The Potential of Natural Propolis Extract Combined with Bovine Bone Graft in Increasing Heat Shock Protein 70 and Osteocalcin on Socket Preservation.

OBJECTIVE: This study aims to combine natural propolis with bovine bone graft (BBG) as a means of extraction socket preservation after 3 and 7 days toward expression of heat shock protein (HSP) 70 and osteocalcin to regenerate bone. MATERIALS AND METHODS: The Cavia cobaya were divided into eight groups, each consisting of seven samples. Their lower left incisors were extracted and induced with PEG, propolis extract, BBG, and a combination of propolis extract BBG. The research subjects were terminated on days 3 and 7 postextraction. Immunohistochemical and histopathological examinations were subsequently performed to observe HSP 70 expression, osteocalcin expression, osteoblasts, and osteoclasts. STATISTICAL ANALYSIS: Data obtained were then analyzed with one-way analysis of variance (ANOVA) and Tukey's honestly significant difference (HSD) tests. RESULTS: Both the groups with the combination of propolis extract and BBG on days 3 and 7 were found to present the highest number of HSP70 expression, osteocalcin expression, and osteoblast cells as well as the lowest number of osteoclasts. CONCLUSION: Both the groups with the combination of propolis extract and BBG on days 3 and 7 were found to present the highest number of HSP70 expression, osteocalcin expression, and osteoblast cells as well as the lowest number of osteoclasts.

Potential of Distilled Liquid Smoke Derived from Coconut (Cocos nucifera L) Shell for Traumatic Ulcer Healing in Diabetic Rats.

OBJECTIVE: Distilled liquid smoke (DLS) is a result of coconut processing by-product that not only serves as a natural food preservative but also has a promising therapeutic effect. The healing potential of DLS derived from coconut (Cocos nucifera L) shell was investigated on a traumatic ulcer with the diabetic rat. MATERIALS AND METHODS: DLS was analyzed the component by gas chromatograph mass spectrometry. Diabetic condition was induced by alloxan in 55 male Wistar rats. Ten mm of traumatic ulcer was made along the labial fornix incisive inferior after the diabetic condition was confirmed. Then DLS coconut shell, benzydamine hydrochloride, and sterile distilled water were applied topically for 3, 5, and 7 days. The potential healing was evaluated based on the expression of nuclear factor kappa beta (NFκB) and tumor necrosis factor alpha (TNF-α) on macrophages using immunohistochemical staining and the amount of collagen using Masson Trichome staining. The difference between each group was analyzed using one-way analysis of variance. The least significant difference test is used to determine the significant difference (p < 0.05). RESULTS: The major compounds found were phenol (36.6%), 2-methoxyphenol (guaiacol) (25.2%), furfural (17.8%), and 4-ethyl-2-methoxyphenol (3.5%) with 28 other minor constituents. The lowest NFκB and TNF-α expression on macrophage was observed by topical application of DLS derived from coconut shell for 3, 5, and 7 days of treatment. The amount of collagen was increased and indicated by the highest result of DLS compared to others. CONCLUSION: The DLS derived from coconut (Cocos nucifera L) shell was able to improve traumatic ulcer healing in a person with diabetes.

High-Mobility Group Box 1 Expression in Mandibular Bone Cells of Experimental Periodontitis.

BACKGROUND: High-mobility group box 1 (HMGB1) was suggested to be associated with the pathogenesis of chronic periodontitis which characterized by alveolar bone loss. HMGB1 was defined as a bone-active cytokine, but the rule of HMGB1 in bone loss of chronic periodontitis is still understood. AIM: The aim of this study is to investigate the expression of HMGB1 on osteoblasts and osteoclasts in the mandible of chronic periodontitis. METHODS: This experimental study was conducted to rats injected by Porphyromonas gingivalis into the buccal and lingual subgingival area at a concentration of 2 × 109 CFU/mL three times a week with 2-day apart for 2, 3, 4, and 6 weeks as chronic periodontitis group and injected by normal saline as control group. Analysis of variance was used to examine the differences between groups followed by least significant difference post hoc test with the level of significance was <0.05. RESULTS: The HMGB1 expression was found in both osteoclasts and osteoblasts of mandibular bone by immunohistochemistry analysis. There was a difference of HMGB1 expression on osteoblasts and osteoclasts of chronic periodontitis. HMGB1 expression was found increased significantly in mandibular osteoblasts of chronic periodontitis, whereas the HMGB1 expression in mandibular osteoclast is higher in 2 and 3 weeks, but it was lower in 4 and 6 weeks. CONCLUSIONS: This study indicated a potential role for HMGB1 in bone loss of chronic periodontitis. HMGB1 on mandibular osteoclasts and osteoblasts may play different rules in the onset and progression of chronic periodontitis.

Effect of Topical Epigallocatechin-Gallate on Lipopolysaccharide-induced Pulpal Inflammation in Rat Models.

Introduction: Pulpal inflammation can be marked by an increase in tumor necrosis factor-α (TNF- α), malondialdehyde (MDA) and calcitonin gene-related peptide (CGRP) level. Epigallocatechin-3-gallate (EGCG) demonstrates the ability to reduce cytokine expression, influence immune receptors, reduce inflammation, neutralize reactive oxygen species (ROS) and to inhibit pain conduction. The present research aimed to determine the anti-inflammatory, antioxidant and pain conduction inhibition of topical EGCG hydrogels in Lipopolysaccharide (LPS)-induced pulpal inflammation in rats. Methods and Materials: A total of 28 male Wistar rats were divided equally into four groups. The negative control group (N) received no treatment, while the positive control group (C) and the other two treatment groups (T1, T2) were induced with LPS for 6 h, followed by the application of topical polyethylene glycol (PEG) hydrogels for C group, 25 ppm EGCG hydrogels for T1 group and 75 ppm EGCG hydrogels for T2 group, before being filled with glass ionomer cement (GIC). After 24 h, PEG and EGCG were reapplied and refilled with GIC for 24 h. The pulp tissue samples were examined by means of immunohistochemistry (IHC) to identify TNF-α, MDA and CGRP expression. All the data obtained was analyzed with one-way analyses of variance (ANOVA) test. Results: The T1 and T2 groups showed a significant decrease in TNF-α and CGRP expression compared to the control group, but there was no significant decrease in MDA in either group (P<0.05). Conclusion: Based on the results of this study, topical application of 75 ppm EGCG hydrogels to the tooth cavities with six hours of pulpal inflammation has the optimal result in reducing the expression of TNF-α and CGRP, but not of MDA.

TLR2 Signaling Pathway in Alveolar Bone Osteogenesis Induced by Aloe vera and Xenograft (XCB)

Abstract The aim of this study was to find the role of TLR2 signaling pathway in reducing osteoclast activity and promoting osteoblast growth by inducing a combination of Aloe vera and cancellous bovine xenograft (XCB) into dental extraction socket. Forty-eight Cavia cobayas were used. They were divided into eight groups (n=6). For control group, their mandibular incisors were extracted and filled with PEG. For treatment groups, they were extracted and filled with XCB, Aloe vera and the combination of Aloe vera and XCB. The first four groups were sacrificed after 7 days and the other groups after 30 days. Immunohistochemistry and histopathology examination were conducted to examine TLR2, TNFa, OPG, collagen-1, and the osteoblast and osteoclast expressions. The expressions of TLR2, OPG and Collagen-1, as well as the number of osteoblast were increased. Meanwhile, the expressions of TNFa and osteoclast were decreased. The study finding was that TLR2 signaling pathway influenced alveolar bone osteogenesis process by reducing osteoclast activity and stimulating osteoblast growth induced by the combination of Aloe vera and XCB.

Resumo O objetivo deste estudo foi investigar o papel da via de sinalização de TLR2 na redução da atividade osteoclástica e na promoção do crescimento de osteoblastos, induzindo uma combinação de Aloe vera e enxerto de osso esponjoso bovino (EOEB) em alvéolo de extração dentária. Quarenta e oito Cavia cobayas foram utilizados e divididos em 8 grupos (n = 6). Para o grupo de controle, seus incisivos mandibulares foram extraídos e preenchidos com polietilenoglicol (PEG). Para grupos de tratamento, os dentes foram extraídos e preenchidos com EOEB, Aloe vera e a combinação de Aloe vera e EOEB. Os primeiros quatro grupos foram sacrificados após 7 dias e os outros grupos após 30 dias. As análises de imunohistoquímica e histopatologia foram realizada para examinar TLR2, TNFa OPG, colágeno-1 e as expressões de osteoblastos e osteoclastos. Houve maior expressão de TLR2, FGF2, OPG e colágeno-1, bem como maior número de osteoblastos. Enquanto isso, a expressão de TNFa e osteoclastos estava diminuída. O principal achado do estudo foi que a via de sinalização de TLR2 influenciou o processo de osteogênese do osso alveolar, reduzindo a atividade dos osteoclastos e estimulando o crescimento de osteoblastos induzido pela combinação de Aloe vera e EOEB.

TLR2 Signaling Pathway in Alveolar Bone Osteogenesis Induced by Aloe vera and Xenograft (XCB).

The aim of this study was to find the role of TLR2 signaling pathway in reducing osteoclast activity and promoting osteoblast growth by inducing a combination of Aloe vera and cancellous bovine xenograft (XCB) into dental extraction socket. Forty-eight Cavia cobayas were used. They were divided into eight groups (n=6). For control group, their mandibular incisors were extracted and filled with PEG. For treatment groups, they were extracted and filled with XCB, Aloe vera and the combination of Aloe vera and XCB. The first four groups were sacrificed after 7 days and the other groups after 30 days. Immunohistochemistry and histopathology examination were conducted to examine TLR2, TNFa, OPG, collagen-1, and the osteoblast and osteoclast expressions. The expressions of TLR2, OPG and Collagen-1, as well as the number of osteoblast were increased. Meanwhile, the expressions of TNFa and osteoclast were decreased. The study finding was that TLR2 signaling pathway influenced alveolar bone osteogenesis process by reducing osteoclast activity and stimulating osteoblast growth induced by the combination of Aloe vera and XCB.

HIV-1 transmitted drug resistance mutations among antiretroviral therapy-Naïve individuals in Surabaya, Indonesia.

BACKGROUND: The emergence of transmitted drug resistance (TDR) compromises the effect of antiretroviral therapy (ART), resulting in treatment failure of human immunodeficiency virus (HIV) disease. Although more than a decade has passed since ART was introduced into Indonesia, information on TDR is limited. Here, a genotypic study of TDR among ART-naïve individuals was conducted in Surabaya, Indonesia. METHOD: HIV-1 seropositive participants were recruited from the communities of commercial sex workers and intravenous drug users as well as from the university teaching hospital in Surabaya. Protease (PR) and reverse transcriptase (RT) genes were sequenced in order to conduct HIV-1 subtyping and phylogenetic analysis and to detect TDR. TDR was defined as the presence of at least one surveillance drug resistance mutation on the WHO list or major drug resistance mutations in the International AIDS Society-USA panel. RESULT: Fifty two and 47 of the PR and RT genes, respectively, were successfully sequenced in the 58 samples. HIV-1 subtyping revealed that 86.3% (50/58) of the sequenced samples were classified as CRF01_AE, 8.6% as subtype B, 3.4% as B/CRF01_AE, and 1.7% as A/G/CRF01_AE. TDR of PR inhibitors was not detected in this study. In contrast, TDR of RT inhibitors was detected in 4.3% (2/47) of samples. In addition, minor drug resistance mutations were detected in 98.1% (51/52) and 12.8% (6/47) of PR and RT genes, respectively. CONCLUSION: This study clarified the predominance of the CRF01_AE strain in Surabaya, Indonesia. The prevalence of TDR was below 5%, indicating that the currently available first-line regimen is still effective in Surabaya. However, the prevalence might be underestimated since we detected only major population of HIV-1 in individuals. Therefore, continuous surveillance is required in order to detect the emergence of TDR in the early phase.

Detection of drug resistance-associated mutations in human immunodeficiency virus type 1 integrase derived from drug-naive individuals in Surabaya, Indonesia.

Although human immunodeficiency virus type 1 (HIV-1) infection causes serious health problems in Indonesia, information in regard to drug resistance is limited. We performed a genotypic study on HIV-1 integrase derived from drug-naive individuals in Surabaya, Indonesia. Sequencing analysis revealed that no primary mutations associated with drug resistance to integrase inhibitors were detected; however, secondary mutations, V72I, L74I/M, V165I, V201I, I203M, and S230N, were detected in more than 5% of samples. In addition, V201I was conserved among all samples. Most integrase genes were classified into CRF01_AE genes. Interestingly, 40% of the CRF01_AE genes had an unusual insertion in the C-terminus of integrase. These mutations and insertions were considered natural polymorphisms since these mutations coincided with previous reports, and integrase inhibitors have not been used in Indonesia. Our results indicated that further studies may be required to assess the impact of these mutations on integrase inhibitors prior to their introduction into Indonesia.

High prevalence of HIV-1 CRF01_AE viruses among female commercial sex workers residing in Surabaya, Indonesia.

BACKGROUND: Human immunodeficiency virus (HIV) infection and acquired immune deficiency syndrome (AIDS) cause serious health problems and have an impact on the Indonesian economy. In addition, the rapid epidemic growth of HIV is continuing in Indonesia. Commercial sex plays a significant role in the spread of HIV; therefore, in order to reveal the current HIV prevalence rate among commercial sex workers (CSWs), we conducted an epidemiological study on HIV infection among CSWs residing in Surabaya, the capital of East Java province of Indonesia with large communities of CSWs. METHODOLOGY/PRINCIPAL FINDINGS: The prevalence of HIV infection among 200 CSWs was studied. In addition, the subtype of HIV type 1 (HIV-1) and the prevalence of other blood-borne viruses, hepatitis B virus (HBV), hepatitis C virus (HCV) and GB virus C (GBV-C), were studied. The prevalence rates of HIV, hepatitis B core antibody, hepatitis B surface antigen, anti-HCV antibodies and anti-GBV-C antibodies were 11%, 64%, 4%, 0.5% and 0% among CSWs involved in this study, respectively. HIV-1 CRF01_AE viral gene fragments were detected in most HIV-positive samples. In addition, most CSWs showed low awareness of sexually transmitted diseases and had unprotected sex with their clients. CONCLUSIONS/SIGNIFICANCE: The HIV prevalence rate among CSWs was significantly higher than that among the general population in Indonesia (0.2-0.4%). In addition, CSWs were at a high risk of exposure to HBV, although chronic HBV infection was less frequently established. Our results suggest the necessity of efficient prevention programs for HIV and other blood-borne viral infections among CSWs in Surabaya, Indonesia.