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"Tujuan dari penelitian ini adalah untuk menganalisis pengaruh Platelet-Rich Plasma (PRP) Eksosom terhadap potensi regenerasi jaringan pulpa gigi dengan evaluasiin vitro viabilitas sel, aktivitas migrasi, dan ekspresi Vascular Endothelial Growth Factor-A (VEGF-A) sel punca pulpa gigi manusia (hDPSCs). HDPSC diambil darisembilan gigi molar tiga dari sembilan donor sesuai kriterian inklusi, dan isolasi serta kultur dilakukan dengan metode enzyme digestion (ED) yang dipanen antara P3 dan P4. Setelah starvatation, hDPSCs dikultur di dalam enam media, yaitu sebagai berikut: Dulbecco's Modified Eagle Medium (DMEM) dan 10% PRP sebagai kelompok kontrol, dan 0,5%, 1%, dan 5% PRP eksosom sebagai kelompok eksperimental. Semua kelompok memiliki tiga rangkap biologis (Triplo). Uji viabilitas sel dievaluasi dengan MTT assay, aktivitas migrasi sel dengan Scratch Assay dan Transwell Migration Assay, dan ekspresi VEGF-A dengan Enzyme- Linked Lmmunosorbent Assay (ELISA). Analisis data dilakukan dengan uji One Way ANOVA (p <0,05) serta uji Kruskal-Wallis dan post hoc Mann-Whitney (p<0,05). Nilai rata-rata viabilitas hDPSCs tertinggi pada 24, 48 dan 72 jam observasi pada kelompok PRP-Eksosom 5% (p <0,05). PRP Eksosom 5% menunjukkan aktivitas migrasi yang lebih tinggi dibandingkan dengan kelompok lain, meskipun tidak terdapat perbedaan bermakna dengan kontrol PRP 10% (p> 0,05). Ekspresi VEGF-A hDPSCs tertinggi terdapat pada kelompok PRP Eksosom 5% pada 72 jampengamatan. Dapat disimpulkan bahwa eksosom PRP 5% berpotensi menginduksi regenerasi pupa gigi manusia.

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The purpose of this study was to analyze the effect of Exosome Platelet-Rich Plasma (PRP) on their potential for human Dental Pulp regeneration by evaluating in vitro cell viability, migration activity, and expression of Vascular Endothelial Growth Factor-A (VEGF-A) of human Dental Pulp Stem Cells (hDPSCs). hDPSCs was taken from nine third molars from nine donors thatfit to the inclusion criteria of this study, isolation and culture were carried out by the enzyme digestion (EZ) method harvested between P3 and P4. After starvatation,hDPSCs were cultured in six media, namely as follows: Dulbecco's Modified EagleMedium (DMEM) and 10% PRP as the control group, and 0.5%, 1%, and 5% PRP exosomes as experimental groups. All groups had a biological triplication (Triplo). Cell viability test was evaluated by MTT assay, cell migration activity with Scratch Assay and Transwell Migration Assay, and VEGF-A expression by Enzyme- Linked Immunosorbent Assay (ELISA). Data analysis was performed using One Way ANOVA (p <0.05) and Kruskal-Wallis (p <0.05) and Pearson/Spearman Correlation test (p<0.05). The highest mean hDPSCs viability was at 24, 48 and 72 hours of observation in the PRP-exosome group 5% (p <0.05). Exosome PRP 5% showed higher migration activity compared to other groups, although there was no significant difference with PRP control 10% (p> 0.05). The highest expression of VEGF-A hDPSCs was found in the PRP exosome group 5% at 72 hours of observation. It can be concluded that the PRP 5% exosome have the highest potential ability in inducing pulp regeneration."

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Tujuan: Menganalisis pengaruh paparan larutan Sodium hipoklorit (NaOCl) dengan konsentrasi 2,5% dan 5,25% pada profil MMP-9 dan struktur kolagen di dentin terhadap kekuatan ikat geser resin komposit-dentin. Metode: Seratus empat puluh empat spesimen dentin dirandom untuk analisis profil MMP-9 dengan pemeriksaan imunohistokimia (n=18) dan ELISA (n=30); analisis struktur kolagen dengan SEM di permukaan oklusal (n=18) dan proksimal (n=18) dan pewarnaan Massons trichrome; serta analisis kekuatan ikat geser resin komposit permukaan oklusal (n=30) dan proksimal (n=30). Spesimen dibagi menjadi 3 kelompok dalam tiap analisis, yaitu: kelompok kontrol, kelompok paparan NaOCl 2,5% dan 5,25%. Hasil: Pemeriksan profil MMP-9 menunjukkan bahwa peningkatan konsentrasi NaOCl dapat menekan profil MMP-9 (p<0,05). Pemeriksaan struktur kolagen menunjukkan bahwa peningkatan konsentrasi NaOCl mampu mendegradasi kolagen dentin (p<0,05). Sodium hipoklorit 5,25% paling efektif menekan jumlah profil MMP-9 (9,9+3,63 ng/mL) dan mendegradasi kolagen serta memiliki nilai kekuatan ikat geser yang paling tinggi (15,85+0,43 MPa) dari pada NaOCl 2,5% (14,51+3,66 ng/mL) dan kelompok kontrol (24,09+8,88 ng/mL; 14,41+0,96 Mpa). Kelompok NaOCl 2,5% memiliki nilai kekuatan ikat geser yang paling rendah (9,2+0,65 MPa). Kesimpulan: Larutan NaOCl 5,25% dapat menekan profil MMP-9 dan mendegradasi kolagen fibril untuk meningkatkan nilai kekuatan ikat geser resin komposit-dentin dan menciptakan suatu ikatan mikro mekanis antara resin dan permukaan anorganik dentin tanpa hybrid layer. 

 Kata kunci: Sodium hipoklorit, profil MMP-9, collagen, dentin, kekuatan ikat geser.

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Objective: To analyze the effect of Sodium hypochlorite (NaOCl) 2.5% and 5.25% exposure on MMP-9 profile and dentin collagen structure toward resin composite-dentin shear bond strength. Method: One hundred and forty four dentin specimens were randomized  for MMP-9 profile analysis using immunohistochemistry staining (n=6) and ELISA (n=10); collagen structure analysis with SEM and Massons trichrome staining (n=6); and resin composite shear bond strength analysis (n=10). Then, specimens were divided into three groups: control, NaOCl 2.5% and 5.25% groups. Results: MMP-9 profile analysis showed that NaOCl concentration increase can suppress MMP-9 profile (p<0.05). Collagen structure analysis showed that NaOCl concentration increase can degrade dentin collagen (p<0.05). NaOCl 5.25% is the most effective in suppressing MMP-9 profile amount (9.9+3.63 ng/mL) and degrading collagen, it also has the highest shear bond strength (15.85 +0.43 MPa) compared to NaOCl 2.5% (14.51+3.66 ng/mL) and control group (24.09+8.88 ng/mL; 14.41+0.96 MPa). NaOCl 2.5% group has the lowest shear bond strength (9.2+0.65MPa). Conclusion: NaOCl 5.25% can suppress MMP-9 profile and degrade fibril collagen to increase the shear bond strengths value and create a micro mechanical bonding between resin and anorganic part of dentin without hybrid layer.

  

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"Latar Belakang: Siler biokeramik berbasis kalsium silikat diketahui memiliki biokompabilitas tinggi, mampu beradhesi dengan dinding saluran akar serta dapat menginduksi respon osteogenik, yang dikategorikan sebagai material biomimetik. Akan tetapi, siler biokeramik memiliki kekurangan pada sifat fisiknya. Kitosan larut air merupakan bentuk modifikasi kitosan yang memiliki keunggulan sebagai antioksidan, antibakteri, berperan dalam penyembuhan lesi dan regenerasi jaringan serta sebagai dapat memperbaiki sifat fisik semen. Perpaduan kedua bahan dapat menciptakan semen saluran akar biomimetik.Tujuan: Melakukan uji karakteristik, sifat fisik dan biologis terhadap novel siler hibrida biokeramik-kitosan larut air (BCC) sebagai semen biomimetik saluran akar.Metode: Novel siler hibrida BCC dimanipulasi dari semen biokeramik (BC) yang telah melalui proses ball-milling dan sintering, kemudian dicampur kitosan larut air dengan W/P 0,4. Empat variabel penelitian, yaitu semen biokeramik BC, novel siler hibrida BCC, siler biokeramik dan siler epoksi resin dilakukan uji karakterisasi (XRD, SEM/EDS), uji sifat fisik (setting time, daya alir, film thickness), serta uji biologis (sitotoksisitas dan bioaktivitas terhadap hPDLSCs).Hasil: Novel siler hibrida BCC mengandung unsur dan puncak kristalin yang serupa dengan siler biokeramik (Sure-Seal Root™), memiliki bentuk partikel yang cenderung globular dan homogen dengan jarak antarpartikel lebih rapat serta ukuran partikel deskriptif yang lebih besar dibandingkan Sure-Seal Root™ dan AH Plus® tetapi lebih kecil dibandingkan semen biokeramik BC. Hanya semen biokeramik BC dan novel siler BCC yang memiliki rasio Ca/Si/P. Novel siler BCC menunjukkan perbedaan setting time, daya alir dan film thickness yang bermakna dengan semen biokeramik BC, Sure-Seal Root™ dan AH Plus®. Keempat kelompok menunjukkan sitotoksisitas rendah terhadap hPDLSCs. Bioaktivitas novel siler BCC relatif lebih tinggi dibandingkan semua kelompok perlakuan dan kontrol.Kesimpulan: Novel siler hibrida BCC berpotensi memberikan implikasi klinis yang baik, menunjukkan sifat fisik yang mendekati standar semen saluran akar dan memiliki bioaktivitas sebagai semen biomimetik saluran akar.

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Background: Currently, bioceramic sealers with calcium silicate based have been developed and show high biocompatibility, are able to adhere to root canal dentin and can induce an osteogenic response, which can be categorized as biomimetic materials. On the other hand, water-soluble particle (WSP) chitosan, one of chitosan derivatives, has advantages as an antioxidant, antibacterial, plays a role in wound healing and tissue regeneration as well as a thickening agent. The combination of these two materials can create a biomimetic endodontic sealer.

Objective: To examine the characteristics, physical and biological properties of novel bioceramic-chitosan hybrid sealer (BCC) as biomimetic endodontic sealer.

Methods: Novel BCC sealer were manipulated from bioceramic cement (BC) which had been synthesized through a ball-milling and sintering process, then mixed with WSP chitosan with a W/P of 0,4. Four variables, namely novel BCC sealer, BC bioceramic cement, bioceramic and epoxy resin sealer were tested characterization (XRD, SEM/EDS), physical properties (setting time, flow, film thickness), and biological tests (cytotoxicity and bioactivity on hPDLSCs).

Results: Novel BCC sealer showed bioceramic elements and crystalline peaks similar to bioceramic sealer (Sure-Seal Root™), had a particle shape that tends to be globular and homogeneous with descriptive particle size larger than Sure-Seal Root™ and AH Plus® but smaller compared to BC bioceramic cement. Only BC bioceramic cements and novel BCC sealer had Ca/Si/P ratios. Novel BCC sealer showed significant differences in setting time, flow, and film thickness with BC bioceramic cements, Sure-Seal Root™ and AH Plus®. The four groups showed low cytotoxicity to hPDLSCs. The bioactivity of novel BCC sealer was relatively higher among other groups and control group.

Conclusion: Novel BCC hybrid sealer has good clinical implications, exhibit physical properties close to standard root canal cements and have bioactivity as root canal biomimetic sealer."

"Latar Belakang : Remineralisasi Guided Tissue Remineralization (GTR) dentin digunakan untuk meningkatkan pembentukan kristal nano ke daerah gap zone serta membangun struktur mineral apatit pada kolagen demineralized dentin dan Carboxymethyl Chitosan (CMC) berperan sebagai protein analog serta memicu remineralisasi dentin. Tujuan: Mendapatkan novel semen carboxymethyl chitosan/amporphous calcium phosphate sebagai agen GTR dentin. Metode: Modifikasi CMC menjadi bubuk CMC/ACP (CA) melalui proses Freeze Dry dan modifikasi bubuk CA melalui proses milling 30 menit menjadi bubuk CMC/ACP paska milling (CAM) dan masing-masing dievaluasi menggunakan FTIR menganalisis gugus fungsi CMC dan gugus fungsi awal CA yang terbentuk. Bubuk CA dan CAM kemudian dicampurkan dengan gipsum hemihydrate pada rasio 5% dan 10% didapatkan kelompok novel semen CAG 5%, CAG 10%, CAMG 5% dan CAMG 10%. Kelompok tersebut dievaluasi menggunakan FTIR, XRD, SEM, setting time, ketahanan kompresi dan MTT assay. Novel semen tersebut diaplikasikan pada dentin terdemineralisasi selama 14 hari dan dievaluasi menggunakan TEM. Hasil: Gugus fungsi CMC berupa -CH2COOH dan N-H terlihat pada bubuk CA dan setelah dilakukan milling menjadi bubuk CAM. Tambahan gugus fungsi fosfat (-PO4) terlihat juga pada bubuk CA dan CAM. Gugus fungsi awal bubuk CA tetap terlihat pada bubuk CAM. Gugus fungsi fosfat (-PO4) terlihat juga pada kelompok novel semen CAG dan CAMG baik 5% dan 10%. Kombinasi bubuk CA dan CAM menggunakan gipsum mempengaruhi fasa mineral material. Kesan topografi berbeda pada novel semen CAG (5% dan 10%) dan novel semen CAMG (5% dan 10%). Terdapat perbedaan bermakna antara kelompok novel semen (CAG 5%, CAG 10%, CAMG 5% dan CAMG 10%) dibandingkan dengan gipsum (kontrol) pada setting time dan ketahanan kompresi (p<0.05). Viabilitas sel dari uji MTT assay menunjukkan novel semen CAMG 10% tidak toksik terhadap hDPSC. Novel semen CAG (5% dan 10%) dan novel semen CAMG (5% dan 10%) menunjukkan kesan remineralisasi dentin ekstrafibrillar dan intrafibrillar pada dentin terdemineralisasi. Kesimpulan: Modifikasi carboxymethyl chitosan melalui proses freeze dry dan milling serta pencampuran rasio 5% dan 10% menggunakan gipsum memiliki kemampuan untuk menginisiasi guided tissue remineralisazation dentin baik secara ekstrafibrillar dan intrafibrillar pada dentin terdemineralisasi.

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Background: Guided Tissue Remineralization (GTR) dentin is used to increase the formation of nanocrystals in the gap zone area and build an apatite mineral structure in demineralized dentin collagen and Carboxymethyl Chitosan (CMC) which acts as a protein analog and triggers dentin remineralization. Objective: To obtain a novel carboxymethyl chitosan/amorphous calcium phosphate cement as a dentinal GTR agent.Purpose: Obtained novel cement carboxymethyl chitosan/amporphous calcium phosphate as an agent for GTR. Methods: Modification of CMC into CMC/ACP (CA) powder through the Freeze Dry process and modification of CA powder through a 30-minute milling process into CMC/ACP powder after milling (CAM) and evaluated using FTIR, respectively, analyzing the CMC functional group and the initial CA functional group formed. CA and CAM powder were then mixed with gypsum hemihydrate at a ratio of 5% and 10% to obtain novel cement groups of CAG 5%, CAG 10%, CAMG 5%, and CAMG 10%. The groups were evaluated using FTIR, XRD, SEM, setting time, compression resistance, and MTT assay. The novel cement was applied to demineralized dentin for 14 days and evaluated using TEM. Result: The CMC functional group (-CH2COOH and N-H) was seen in CA and CAM powder. Addition of phosphate (-PO4) functional grup were detected in CA and CAM powder. Phosphate (-PO4) functional group was seen in novel cement CAG (5% and 10%) and CAMG (5% and 10%). The combination of gipsum using CA and CAM powders produces different mineral phases. Topographical impressions differed between Novel cement groups of CAG (5% and 10%) and CAMG (5% and 10%). There was a significant difference between the novel cement groups (CAG 5%, CAG 10%, CAMG 5%, and CAMG 10%) compared to gypsum (control) in setting time and compression resistance (p<0.05). Viability cell confirmation using MTT assay showed that novel cemen group of CAMG 10% did not toxic to hDPSC. Novel cement groups of CAG (5% and 10%) and CAMG (5% and 10%) demonstrated the effects of extrafibrillar and intrafibrillar dentin remineralization on demineralized dentin. Conclusion: Modifying carboxymethyl chitosan through freeze dry and milling processes and modifying the mixing ratio of 5% and 10% using gypsum can initiate guided tissue remineralization of dentin both extrafibrillarly and intrafibrillarly in demineralized dentin."

"Latar Belakang: pulpa memiliki sifat low-compliance yang memengaruhi proses regenerasinya. Tujuan: menganalisis potensi Platelet- Rich Plasma (PRP) Eksosom terhadap regenerasi pulpa gigi secara in-vitro viabilitas sel, aktivitas migrasi, dan ekspresi Vascular Endothelial Growth Factor-A (VEGF-A) hDPSCs. Metodologi: hDPSC sembilan gigi molar tiga dikultur dengan metode enzyme digestion (ED) yang dipanen pada P3 dan P4. Kemudian dikultur di dalam enam media, yaitu: Dulbecco's Modified Eagle Medium (DMEM) dan 10% PRP sebagai kelompok kontrol, dan 0,5%, 1%, dan 5% eksosom dari PRP. Semua kelompok memiliki tiga rangkap biologis (Triplo). Uji viabilitas sel dievaluasi dengan MTT assay, aktivitas migrasi sel dengan Scratch Assay dan Transwell Migration Assay, dan ekspresi VEGF-A dengan Enzyme-Linked Lmmunosorbent Assay (ELISA). Analisis data dilakukan dengan uji One Way ANOVA (p <0,05) serta uji Kruskal-Wallis dan post hoc Mann-Whitney (p <0,05). Hasil: viabilitas hDPSCs tertinggi pada 24, 48 dan 72 jam observasi pada kelompok Eksosom dari PRP 5% (p <0,05). Eksosom dari PRP 5% menunjukkan aktivitas migrasi yang lebih tinggi dibandingkan dengan kelompok lain, meskipun terdapat perbedaan tidak bermakna dengan kontrol PRP 10% (p >0,05). Ekspresi VEGF-A hDPSCs tertinggi terdapat pada kelompok PRP Eksosom 5% pada 72 jam observasi. Kesimpulan: eksosom dari PRP 5% berpotensi menginduksi regenerasi pupa gigi manusia.

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Background: pulp has low-compliance properties that affect its regeneration process. Objective: to analyze the potential of Platelet-Rich Plasma (PRP) exosomes on the regeneration of dental pulp by in-vitro evaluation of cell viability, migration activity, and expression of Vascular Endothelial Growth Factor-A (VEGF-A) hDPSCs. Methodology: hDPSCs of nine third molars cultured by enzyme digestion (ED) method were harvested at P3 and P4. Then cultured in six media, Dulbecco's Modified Eagle Medium (DMEM) and 10% PRP as a control group, and 0.5%, 1%, and 5% exosomes of PRP. All groups had a biological triple (Triplo). Cell viability assay was evaluated by MTT assay, cell migration activity by Scratch Assay and Transwell Migration Assay, and VEGF-A expression by Enzyme-Linked Lmmunosorbent Assay (ELISA). Data analysis was performed using One Way ANOVA (p < 0.05) and Kruskal-Wallis and post hoc Mann-Whitney tests (p < 0.05). Results: The viability of hDPSCs was highest at 24, 48 and 72 hours of observation in the Exosomes group of 5% PRP (p < 0.05). Exosomes from 5% PRP showed higher migratory activity compared to other groups, although there was no significant difference with 10% PRP control (p > 0.05). The highest expression of VEGF-A hDPSCs was found in the 5% PRP Exosomes group at 72 hours of observation. Conclusion: exosomes of 5% PRP have the potential to induce the regeneration of human dental pulp."