POLA EKSPRESI PVRL1 DAN IRF6 PADA PENDERITA CLEFT LIP AND PALATE

Latar Belakang: Cleft Lip and Palate (CLP) merupakan kelainan kongenital berupa celah yang melibatkan bibir, tulang alveolar, palatum, dan septum nasal. Penderita CLP mengalami gangguan estetik dan fungsional. Kelainan CLP merupakan dampak dari gangguan proses palatogenesis pada masa embrionik. Etiologi dari CLP dipengaruhi oleh faktor genetik dan lingkungan. Kejadian CLP terkait dengan defek pada IRF6 dan PVRL1 Tujuan: Untuk menganalisis perbedaan profil protein total pada penderita CLP, menganalisis perbedaan nilai Rf, serta mengidentifikasi target protein IRF6 dan PVRL1. Metode: Dilakukan teknik SDS-PAGE dengan konsentrasi gel 12% menggunakan plasma dari penderita CLP, orang tua penderita, dan kontrol. Setelah proses elektroforesis, dilakukan proses staining menggunakan Coomassie Blue. Selanjutnya dilakukan uji dot-blot untuk identifikasi protein target IRF6 dan PVRL1. Hasil: Didapatkan hasil uji positif terhadap protein IRF6 dan PVRL1 melalui uji dotblot. Terdapat perbedaan nilai Rf yang signifikan pada pita A dan D. Didapatkan intensitas protein dengan kisaran berat molekul 15-20 kDa, 45-60 kDa yang diduga sebagai target protein IRF6, dan 70-100 kDa yang diduga sebagai target protein 70-100 kDa yang lebih tinggi pada kelompok penderita CLP dibandingkan kelompok kontrol. Namun tidak terdapat perbedaan intensitas yang signifikan antar penderita CLP dengan orang tua. Kesimpulan: Terdapat perbedaan ekspresi profil protein yang signifikan antara penderita CLP daripada kelompok kontrol

The amount of macrophages and activated plasma cells on wound healing process affected by spirulina

Background: Spirulina which grows abundantly in tropical seas have been investigated to enhance immune system. The administration of spirulina in tooth extraction sockets was expected to optimise the function of immunocompetent cells. Therefore, wound healing process would be improved. Purpose: The aim of this study was to prove that administration of spirulina could influence immune system in tooth extraction sockets. Method: There were 28 Cavia cobayas used in this study and were put in group of four. Mandibular left incisive were extracted from each of them. The basis made from mixture of polyethylene glycol (PEG) 400 and PEG 4000 was administrated into each socket in control group (TG0). In addition, spirulina 12% was administrated into group TG1, spirulina 24% was administrated into group TG2, and spirulina 48% was administrated into group TG3. All of the Cavia cobaya were decapitated and the jaws were removed in day 5 after tooth extraction. The jaws were decalcified in EDTA solution, formed into paraffin block, processed for hematoxylin and eosin (H & E) and immunohistochemistry staining afterwards. Datas were analysed statistically using Anova method. Result: There was an augmentation in the number of macrophages and activated plasma cells after spirulina application. The administration of higher concentrations of Spirulina leads to greater amount of macrophages and activated plasma cells in each groups. Conclusion: In conclusion, spirulina is able to increase the amount of macrophages and activated plasma cells which play important role in healing process

Soluble Human Leukocyte Antigen Molecules Detected in Orofacial Cleft Patients: A Case-Control Study

Objective: To compare soluble HLA-C and HLA-DR molecules present in the plasma of orofacial cleft and non-orofacial cleft populations. Material and Methods: Orofacial cleft patients were recruited using an accidental sampling approach (n=15). Peripheral blood was collected from the participants and processed for Enzyme Linked Immunosorbent Assay (ELISA) against HLA-C and HLA-DR with specific antibodies. The absorbance was calculated utilizing ELISA reader. Data were statistically analyzed using an independent t-test to compare the disease and control groups. Results: The levels of soluble HLA-C and HLA-DR were significantly higher in the diseased group compared to the control group (p<0.05). Conclusion: The role of HLA molecules in non-communicable disease and congenital anomalies, particularly orofacial cleft, remains speculative despite the positive results of this study and  those  of previous investigations. It suggests that the variables examined may affect specific pathways involved in the pathogenesis of orofacial cleft, and predispose the individuals concerned to the oral cleft

The immunomodulatory effect of green tea (Camellia sinensis) leaves extract on immunocompromised Wistar rats infected by Candida albicans

Background and Aim: The immunocompromised condition is considered a defect in the immune system. This condition tends to increase the risk of oral candidiasis, due to the inability of the immune system to eliminate the adhesion of Candida albicans and leads to systemic candidiasis with a mortality rate of 60%. Green tea (Camellia sinensis) contains potential antioxidant and immunomodulatory which acts as anticancer, antifungal, and antivirus agent. The aim of this study was to invent herbal-based medicine, which acts as an immunomodulator and antifungal agent to treat fungal infection in immunocompromised patients. Materials and Methods: Thirty-five immunocompromised Wistar rats induced with C. albicans were divided into 7 groups (n=5): Control group (C+); treated for 4 days with green tea extract 1.25% (GT 4), epigallocatechin gallate (EGCG) 1% (EGCG 4), EGC 1% (EGC 4); and treated for 7 days with green tea extract 1.25% (GT 7), EGCG 1% (EGCG 7), and EGC 1% (EGC 7). Tongue tissue was collected and analyzed with immunohistochemistry staining using monoclonal antibody; interleukin (IL)-17A, IL-8, and human beta-defensin 2 (HBD)-2. Data were analyzed using analysis of variance test and Tukey honest significant differences test. Results: The expression of IL-17A, IL-8, and HBD-2 was significantly increased (p=0.000) after green tea extract administration in 7 days, whereas in 7 days, the expression of IL-8, IL-17A, and HBD-2 after EGCG and EGC administration did not give a significant result (p>0.005). Conclusion: Within the limits of this study, green tea extract has the ability as an immunomodulatory agent in an immunocompromised patient infected by C. albicans through expression augmentation of IL-8, IL-17A, and HBD-2 compared to EGCG and EGC

Effect of Vitamin D during Orthodontic Tooth Movement on Receptor Activator of Nuclear Factor Kappa-Â Ligand Expression and Osteoclast Number in Pregnant Wistar Rat (Rattus novergicus)

Background: Female patients have the possibility to become pregnant during orthodontic treatment. Vitamin D usually consumed by pregnant women. Estrogen and Vitamin D could affect bone metabolism. Aim and Objectives: The aim of this study was to analyze the effect of vitamin D during orthodontic movement in pregnant rats by Receptor Activator of Nuclear Factor Kappa-Â Ligand (RANKL) expression and osteoclast number. Material and Methods: The experimental observational analytic study with post-test only control group design and simple random sampling method was conducted. 24-healthy-female Wistar rats were divided into 4 groups; K1: pregnant rats with orthodontic tooth movement and vitamin D on Day 7; K2: pregnant rats with orthodontic tooth movement and vitamin D on Day 14; K3: pregnant rats with orthodontic tooth movement without vitamin D on Day 7 and; K4: pregnant rats with orthodontic tooth movement without vitamin D on Day 14. Nickle-Titanium coil spring with 10 g/mm2 force was placed between the incisors and the maxillary molars. The RANKL expression and osteoclasts number were analyzed using Analysis of Variance (ANOVA) (p<0.05). Results: The highest osteoclasts number (8.494 ± 1.194), and RANKLexpression (7.967 ± 2.185) found in K1 group with significant between groups (p<0.05).Conclusions: Vitamin D increase osteoclast number and RANKL expression during orthodontic tooth movement in pregnant rats

Soluble Human Leukocyte Antigen Molecules Detected in Orofacial Cleft Patients: A Case-Control Study

Objective: To compare soluble HLA-C and HLA-DR molecules present in the plasma of orofacial cleft and non-orofacial cleft populations. Material and Methods: Orofacial cleft patients were recruited using an Accidental sampling approach (n=15). Peripheral blood was collected from the participants and processed for Enzyme Linked Immunosorbent Assay (ELISA) against HLA-C and HLA-DR with specific antibodies. The absorbance was calculated utilizing ELISA reader. Data were statistically analyzed using an independent t-test to compare the disease and control groups. Results: The levels of soluble HLA-C and HLA-DR were significantly higher in the diseased group compared to the control group (p<0.05). Conclusion: The role of HLA molecules in non-communicable disease and congenital anomalies, particularly orofacial cleft, remains speculative despite the positive results of this study and those of previous investigations. It suggests that the variables examined may affect specific pathways involved in the pathogenesis of orofacial cleft, and predispose the individuals concerned to the oral cleft

High Mobility Group Box 1 and Heat Shock Protein-70 Expression Post (-)-Epigallocatechin-3-Gallate in East Java Green Tea Methanolic Extract Administration During Orthodontic Tooth Movement in Wistar Rats

Objective: To investigate the expression of High Mobility Group Box 1 (HMGB1) and Heat Shock Protein-70 (HSP-70) during orthodontic tooth movement (OTM) after (-)- Epigallocatechin-3-Gallate(EGCG) in East Java Green Tea (Camelia Sinensis) Methanolic Extract (GTME) administration in vivo. Material and Methods: 28 Wistar rats (Rattus Novergicus) was used and divided into 4 groups accordingly: K- without EGCG and OTM; K+ with OTM, without EGCG for 14 days; T1with OTM for 14 days and EGCG for 7 days; treatment group 2 (T2) with OTM and EGCG for 14 days. OTM animal model was achieved through the installation of the OTM device by means of NiTi close coil spring with 10g force placed between the first incisor and first maxillary molars. The samples were terminated on Day 14. The pre-maxillary was isolated for the immunohistochemical examination. Analysis of Variance (ANOVA) then continued with Tukey Honest Significant Difference (HSD) (p<0.05) was performed to analyze the data. Results: The highest HMGB1 and HSP-70 expression were found in the K+ group pressure side, meanwhile the lowest HMGB1 and HSP-70 expression were found in K- group tension side in the alveolar bone. There was a significant decrease of HMGB1 and HSP-70 expression in T2 compared to T1 and K+ with significant between groups (p<0.05; p=0.0001). Conclusion: The decreased expression of HMGB1 and HSP-70 in alveolar bone of OTM wistar rats due to post administration of GTME that consisted EGCG

High Mobility Group Box 1 and Heat Shock Protein-70 Expression Post (-)-Epigallocatechin-3-Gallate in East Java Green Tea Methanolic Extract Administration During Orthodontic Tooth Movement in Wistar Rats

Objective: To investigate the expression of High Mobility Group Box 1 (HMGB1) and Heat Shock Protein-70 (HSP-70) during orthodontic tooth movement (OTM) after (-)- Epigallocatechin-3-Gallate (EGCG) in East Java Green Tea (Camelia Sinensis) Methanolic Extract (GTME) administration in vivo. Material and Methods: 28 Wistar rats (Rattus Novergicus) was used and divided into 4 groups accordingly: K- without EGCG and OTM; K+ with OTM, without EGCG for 14 days; T1with OTM for 14 days and EGCG for 7 days; treatment group 2 (T2) with OTM and EGCG for 14 days. OTM animal model was achieved through the installation of the OTM device by means of NiTi close coil spring with 10g force placed between the first incisor and first maxillary molars. The samples were terminated on Day 14. The pre-maxillary was isolated for the immunohistochemical examination. Analysis of Variance (ANOVA) then continued with Tukey Honest Significant Difference (HSD) (p&lt;0.05) was performed to analyze the data. Results: The highest HMGB1 and HSP-70 expression were found in the K+ group pressure&nbsp; side, meanwhile the lowest HMGB1 and HSP-70 expression were found in K- group tension side in the alveolar bone. There was a significant decrease of HMGB1 and HSP-70 expression in T2 compared to T1 and K+ with significant between groups (p&lt;0.05; p=0.0001). Conclusion: The decreased expression of HMGB1 and HSP-70 in alveolar bone of OTM wistar rats due to post administration of GTME that consisted EGCG

Pola Ekspresi PVRL-1 Dan IRF6 Pada Penderita Cleft Lip And Palate

Celah orofasial merupakan anomali kongenital yang sering terjadi dengan etiologi multifaktorial. Pada umumnya kelainan ini dipengaruhi oleh faktor genetik dan lingkunganyang memicu terjadinya defek genetik, dan secara klinis menimbulkan deformitas wajah. Pada data nasional tercatat bahwa prevalensi CLP di Indonesia sebesar 0,2%. Pada tahun 2012organisasi internasional Pusat Pelatihan Celah Bibir menyatakan peningkatan kasus CLP sekitar 7500 kasus per tahun di Indonesia. Penderita CLP yang terdaftar pada Yayasan CLP Surabaya berjumlah 1257 pasien. Cleft Lip and Palate (CLP) paling banyak terjadi pada ras Asia dan keturunan Indian di Amerika, yaitu satu kasus dari 500 kelahiran. Cleft Lip and Palate (CLP) merupakan hasil dari gangguan pembeimikan bibir pada minggu ke-4 sampai ke-7 pada masa embryogenesis. Adanya gangguan pada proses tersebut menyebabkan celah pada bibir dan/atau palatum. Keluhan yang banyak ditemukan pada penderita CLP adalah adanya problem pengunyahan, infeksi telinga yang rekuren, maloklusi dan gangguan fonetik, sehingga dibutuhkan perawatan multidisiplin dari bidang obstetrik, pediatri bedah plastik, rehabilitasi, dan kedokteran gigi. Faktor lingkungan pada masa kehamilan yang berpengaruh pada proses palatogenesis adalah infeksi pada ibu hamil, usia ibu yang lebih dari 40 tahun saat hamil, defisiensi nutrisi, paparan bahan teratogenik saat masa kehamilan, dan kebiasaan merokok, baik aktif maupun pasif. Terjadinya CLP karena factor genetik merupakan akibat dari ekspresi berbagai gen yang diduga memiliki peran terhadap palatogenesis, antara lain: TBX22, PVRL1, dan IRF6, tetapi hasil tersebut dapat berbeda, dipengaruhi oleh lokasi geografis, etnis dan ras. Pada penelitian ini dikembangkan pemeriksaan imunogenetik menggunakan specimen saliva. Saliva merupakan refleksi dari kondisi sistemik tubuh, tidak terbatas pada keadaan di rongga mulut. Penelitian mengenai cleft lip and palate sudah banyak dilakukan di Indonesia, akan tetapi belum memberikan pemahaman yang mendalam tentang pola ekspresi protein dari PVRL,1 dan IRF6. Berdasar hal tersebut maka perlu dilakukan penelitian untuk rnenjawab permasalahan yang mendasari terjadinya CLP di Indonesia

The amount of macrophages and activated plasma cells on wound healing process affected by spirulina

Background: Spirulina which grows abundantly in tropical seas have been investigated to enhance immune system. The administration of spirulina in tooth extraction sockets was expected to optimise the function of immunocompetent cells. Therefore, wound healing process would be improved. Purpose: The aim of this study was to prove that administration of spirulina could influence immune system in tooth extraction sockets. Method: There were 28 Cavia cobayas used in this study and were put in group of four. Mandibular left incisive were extracted from each of them. The basis made from mixture of polyethylene glycol (PEG) 400 and PEG 4000 was administrated into each socket in control group (TG0). In addition, spirulina 12% was administrated into group TG1, spirulina 24% was administrated into group TG2, and spirulina 48% was administrated into group TG3. All of the Cavia cobaya were decapitated and the jaws were removed in day 5 after tooth extraction. The jaws were decalcified in EDTA solution, formed into paraffin block, processed for hematoxylin and eosin (H & E) and immunohistochemistry staining afterwards. Datas were analysed statistically using Anova method. Result: There was an augmentation in the number of macrophages and activated plasma cells after spirulina application. The administration of higher concentrations of Spirulina leads to greater amount of macrophages and activated plasma cells in each groups. Conclusion: In conclusion, spirulina is able to increase the amount of macrophages and activated plasma cells which play important role in healing process