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Abstrak :
Sel punca adiposa (SPA) telah menjadi sumber pengobatan terkini yang menjanjikan. Penelitian terbaru menunjukkan bahwa mekanisme yang mendasari manfaat sel punca sebagai pengobatan berhubungan dengan efek modulasi parakrin daripada pemberian sel punca itu sendiri. Beberapa penelitian tentang SPA telah melaporkan adanya perubahan dalam jumlah, proliferasi, dan potensi diferensiasi, serta adanya penurunan sitokin dan growth factor pada SPA sehubungan dengan usia donor. Penelitian ini bertujuan untuk menyelidiki perubahan sekretom berdasarkan kelompok usia tua dan muda serta dampaknya terhadap sel fibroblas yang diinduksi UVB. Sel punca adiposa dari donor usia tua dan muda dikultur dan diisolasi untuk diambil sekretomnya. Sel fibroblas yang diinduksi UVB kemudian diinkubasi dengan sekretom sel punca adiposa tersebut. Parameter seperti viabilitas sel dan konsentrasi TGF-β1 pada sekretom sel punca adiposa serta presentase apoptosis, sel mati, ekpresi gen MMP-3 dan COL1A pada sel fibroblas yang diinduksi UVB dan diberikan sekretom usia donor berbeda dievaluasi untuk menilai efek perbedaan usia donor tersebut. Hasil penelitian menunjukan bahwa sekretom sel punca adiposa dengan usia donor berbeda memberikan dampak yang berbeda pada sel fibroblas yang diinduksi UVB. Konsentrasi TGF-β1 pada sekretom SPA asal donor usia muda lebih tinggi dibandingkan dengan SPA donor tua. Persentase apoptosis, sel mati dan ekpresi gen MMP-3 terhadap GAPDH lebih tinggi pada sel fibroblas yang diinduksi UVB dan diberi sekretom SPA donor tua dibandingkan dengan yang diberi sekretom SPA donor muda. Ekpresi gen COL1A lebih rendah pada pada sel fibroblas yang diinduksi UVB dan diberikan sekretom SPA donor tua dibandingkan diberikan sekretom SPA donor muda. Oleh sebab itu, dapat disimpulkan adanya penurunan kemampuan sekretom sel punca adiposa usia donor tua dalam memperbaiki penuaan pada sel fibroblas yang dipajan dengan UVB, berdasarkan morfologi sel, presentase sel apoptosis dan sel mati serta ekpresi MMP-3 dan COL1A terhadap GAPDH. ......Adipose stem cells (ASC) have become a promising source of current treatment. Recent research suggests that the mechanisms underlying the benefits of stem cells as a treatment relate to paracrine modulatory effects rather than the administration of stem cells themselves. Several studies on ASC have reported changes in the number, proliferation and differentiation potential, as well as a decrease in cytokines and growth factors in ASC in relation to donor age. This study aims to investigate secretome changes based on young and old age groups and their impact on UVB- induced fibroblast cells. Adipose stem cells from young and old donors were cultured and isolated for secretome collection. The UVB-induced fibroblast cells were then incubated with the secretome of the adipose stem cells. Parameters such as cell viability and TGF-β1 concentration in the secretome of adipose stem cells as well as the percentage of apoptosis, dead cells, MMP-3 and COL1A gene expression in fibroblast cells induced by UVB and given the secretome of different donor ages were evaluated to assess the effect of differences in donor age. The results showed that the secretome of adipose stem cells with different donor ages had different impacts on UVB-induced fibroblast cells. The concentration of TGF-β1 in the secretome of SPA from young donors was higher compared to ASC from old donors. The percentage of apoptosis, cell death and expression of the MMP-3 gene against GAPDH was higher in fibroblast cells induced by UVB and given the SPA secretome of old donors compared to those given the ASC secretome of young donors. COL1A gene expression was lower in fibroblast cells induced by UVB and given the ASC secretome of old donors compared to those given the ASC secretome of young donors. Therefore, it can be concluded that there is a decrease in the ability of the secretome of adipose stem cells from old donors to improve aging in fibroblast cells exposed to UVB, based on cell morphology, the percentage of apoptotic cells and dead cells as well as the expression of MMP-3 and COL1A against GAPDH.

Abstrak :
Peningkatan penggunaan Fetal Bovine Serum (FBS) dari tahun ke tahun menyebabkan permintaan melebihi supply. Hal ini menyebabkan peningkatan harga FBS. Penelitian ini dimulai dengan membuat sediaan serbuk royal jelly dan propolis menggunakan metode freeze drying. Serbuk royal jelly dan ekstrak propolis kemudian akan ditambahkan pada media kultur kemudian dihitung proliferasi dan viabilitas sel fibroblas dengan menggunakan Trypan blue assay. Penambahan variasi propolis, royal jelly, dan FBS akan dilakukan dalam 7 variasi konsentrasi. Hasil proliferasi sel fibroblas media variasi 1 pada hari ketujuh memberikan hasil terbaik dari antara media variasi lain dengan rata-rata 24.780,7±401,98 sel/cm2. Namun hasil tersebut masih belum melebihi proliferasi pada media kontrol. Hasil pengujian viabilitas media variasi 1 hingga media variasi 4 pada hari ketujuh memberikan hasil viabilitas > 95%. Berdasarkan pada hasil PDL, viabilitas, dan morfologi sel fibroblast, maka media variasi terbaik merupakan media variasi 3 dengan waktu inkubasi 3 hari yang mencapai nilai level pembelahan sebanyak 3,3 level. Untuk waktu inkubasi 7 hari, media 1 merupakan yang terbaik diantara media variasi dengan nilai level pembelahan sebanyak 3,4level. ......The increasing use of Fetal Bovine Serum (FBS) over the years has led to a demand exceeding the supply. This has resulted in an increase in the price of FBS. This research begins by preparing powdered formulations of royal jelly and propolis using the freeze-drying method. The powdered royal jelly and powder propolis sulawesi extracts will then be added to the culture media, and the proliferation, viability, and population doubling level (PDL) of sel fibroblas cells will be assessed using the Trypan blue assay. Seven serum concentration variations (combination of royal jelly, propolis, and FBS) will be added on multi well culture plate 12 wells microplate sterile. The results of sel fibroblas cell proliferation in 1st media variation on the seventh day showed the best outcome among the other variations, with an average of 24,780.7±401.98 cells/cm2. The viability testing results of 1st to 4th variation media on the seventh day showed viability rates of >95%. Based on the results of PDL, viability, and fibroblast cell morphology, the best variation medium is variation medium 3 with an incubation time of 3 days, reaching a cell division level of 3.3. For an incubation time of 7 days, variation medium 1 is the best among the 1st variation with seven day incubation, at the average level of 3.38 PDs.

Abstrak :
Kulit merupakan sistem pertahanan eksternal, langsung menjalani proses penyembuhan luka ketika terjadi luka dan banyak orang cenderung memberi proses penyembuhan luka dengan agen antiseptik, povidone iodine 10 Betadine . Namun, terdapat ide baru tentang penggunaan povidone iodine 5 pada penyembuhan luka kulit yang dapat memberikan efek yang berbeda. Tujuan dari penelitian ini adalah untuk membandingkan efek berbeda dari penggunaan konsentrasi yang berbeda dari povidone iodine pada jumlah PMN, fibroblast, dan serat kolagen dan untuk menentukan kadar 5 atau 10 yang lebih cocok untuk digunakan. Penelitian ini menggunakan tikus sebagai sampel, masing-masing tikus diberikan 3 luka dengan 3 perlakuan berbeda terdiri dari kontrol, povidone iodine 10 , dan povidone iodine 5 . Pada hari ke-3, tiga tikus pertama dikorbankan dan pada hari ke-7 3 tikus berikut dikorbankan, lalu dibuat spesimen histologi dengan mengambil area luka dan diwarnai dengan Hematoksilin-Eosin untuk menganalisis jumlah PMN dan fibroblast, serta Van Gieson menganalisis serat kolagen. Hasil dari penelitian ini memperlihatkan tidak ada perbedaan yang signifikan antara povidone iodine 5 dan 10 dalam proses keseluruhan penyembuhan luka yang dilihat dari jumlah PMN, fibroblast, dan serat kolagen. ......Skin is an external defense system, directly undergo wound healing process when scars occur and people tend to interfere the wound healing process with antiseptic agents, in this case is the use of povidone iodine 10 Betadine . However, there is new idea about the appliance of povidone iodine 5 on cutaneous wound healing may give different effect. This research aims to compare the different effect of using different concentration of povidone iodine on number of PMN, fibroblast, and collagen fibers during wound healing process and to determine which one is more suitable to use. This experiment using rats as samples, each rat is given 3 wounds with 3 different treatments consisted of control, povidone iodine 10, and povidone iodine 5. On the 3rd day, the first three rats were sacrificed and on the 7th day the following 3 rats were sacrificed, then made histological specimens by taking the wound area and stained it using Hematoxylin eosin to analyze number of PMN and fibroblast, also Van Gieson to analyze collagen fibers. The result of this experiment is that there is no significant difference among povidone iodine 5 and 10 in overall process or phases of wound healing, as seen from number of PMN, fibroblast, as well as collagen fibers.

Abstrak :
Bacterial lipopolysaccharide (LPS) is impacted in the etiology of inflammatory periodontal disease. Aside from immunopathologic reactions which may be involved in the pathogenesis of the disease, the possibility exist that direct cytotoxic effect on cultured human gingival fibroblasts may be equally destructive. The expression of P53 protein can be one of markers to examine the state of impaired DNA. The purpose of this study was to investigate the effect of LPS toward expression of P53 protein on cultured human gingival fibroblasts. Cultured human gingival fibroblasts were exposed to LPS in concentrations of 50 and 200 ug/ml and untreated medium for a period of 24 and 48 hours. Cells were harvested and prepared for immunohistochemical evaluation. After exposure for 24 and 48 hours, the fraction of P53-positive cells was 81.7% in case of 50 ug/ml LPS, and 88.8% in case of 200 ug/ml LPS. After exposure for 48 hours, the fraction of P53-positive cells was 32.2% in case of 50 ug/ml LPS, and 21.1% in case of 200 ug/ml LPS. None of untreated group showed p53-positive cells. Up-regulation of p53 protein during the initial logarithmic phase of growth may be a consequence of on-going DNA damage.

Abstrak :
Manfaat madu untuk penyembuhan luka sudah banyak diteliti, namun informasi manfaatnya untuk penyembuhan luka timpanoplasti masih terbatas. Penelitian ini bertujuan mengevaluasi efek madu manuka (Mn) dan madu trigona (Tr) asli Indonesia pada re-epitelisasi membran timpani (MT) melalui potensi proliferasi fibroblas, keratinosit, sekresi KGF dan basic-FGF. Penelitian in vivo berupa uji klinis acak, tersamar ganda, pada 64 pasien dewasa otitis media supuratif kronik (OMSK) tipe aman tenang yang menjalani timpanoplasti di RSUPN dr. Cipto Mangunkusumo pada bulan Juni 2021–Agustus 2022. Pasien diacak dan disamarkan ke dalam dua kelompok, yaitu diberikan gelfoam plus gel Mn 100% medical grade (intervensi) atau hanya diberikan gelfoam (kontrol) di liang telinga saat timpanoplasti. Tampon telinga diangkat setelah dua minggu dan pasien diminta kontrol setiap minggu selama enam minggu. Penelitian in vitro dilakukan di Laboratorium Universitas YARSI. Kultur fibroblas dan keratinosit yang diisolasi dari pasien OMSK diberikan pajanan Mn dan Tr dengan tiga konsentrasi yaitu 0,04%, 0,1%, dan 0,25%, kemudian dilakukan uji proliferasi, KGF dan bFGF juga diukur dan dibandingkan dengan kelompok kontrol. Proporsi pengeringan luka pascatimpanoplasti kelompok intervensi lebih banyak secara bermakna dibandingkan kontrol pada minggu ke-3, ke-4, dan ke-6. Madu manuka dan Tr tidak meningkatkan jumlah sel kultur fibroblas, tetapi mempersingkat durasi doubling time. Jumlah sel kultur keratinosit lebih tinggi secara bermakna dibandingkan kontrol pada semua kelompok Mn dan Tr 0,04%. Sekresi KGF meningkat seiring pertambahan sel. Pada hari ke-6 dan hari ke-8, sekresi KGF lebih tinggi pada beberapa kelompok intervensi dibandingkan kontrol. Sebaliknya, kadar bFGF menurun seiring pertambahan sel. Terdapat korelasi positif antara lama pajanan kedua jenis madu dengan proliferasi fibroblas. Lama pajanan Mn 0,04%, 0,1%, dan Tr 0,04% berkorelasi positif dengan jumlah sel kultur keratinosit. Disimpulkan pemberian Mn saat timpanoplasti meningkatkan pencapaian re-epitelisasi MT sempurna melalui efeknya pada fibroblas dan keratinosit, serta berpotensi meningkatkan keberhasilan timpanoplasti. Penelitian ini juga menunjukkan efek positif Tr pada fibroblas dan keratinosit, sehingga potensi terapeutik madu ini dapat diteliti lebih lanjut ......Benefits of honey on wound healing have been widely reported, but information about its effect on the re-epithelialization of the tympanic membrane (TM) is limited. This study aims to evaluate the effect of manuka honey (MH) and trigona honey (TH) from Indonesia, on TM re-epithelization through their potential action on the proliferation of fibroblasts, keratinocytes, secretion of KFG and basic-FGF. The in vivo study was a randomized, controlled, double-blind clinical trial on 64 adult patients with mucosal type chronic suppurative otitis media (CSOM) undergoing tympanoplasty at Cipto Mangunkusumo General Hospital from June 2021–August 2022. Patients were randomized and blinded into two groups, receiving either gel foam soaked in 100% medical grade MH (intervention group) gel or only gel foam (control group) placed in the external auditory canal during tympanoplasty. The ear tampon was removed after two weeks, and patients were followed up weekly for six weeks. The in vitro study was conducted at the YARSI University Laboratory. Fibroblast and keratinocyte cultures isolated from CSOM patients were exposed to MH and TH with three dilutions: 0.04%, 0.1%, and 0.25%. The cells were then subjected to proliferation assays, KGF and bFGF were also assessed and compared with the control group. The intervention group had a significantly higher proportion of dry tympanoplasty wounds than control at the 3rd, 4th, and 6th visit. Manuka honey and TH did not increase the number of fibroblasts but shortened the doubling time duration. A significantly higher number of keratinocytes than control was observed in all MH groups and the 0.04% TH group. KGF secretion increased as the number of cells increased. On day 6 and day 8, KGF secretion was higher in some of the intervention groups compared with the control group. In contrast, fibroblast bFGF secretion decreased as the number of cells increased. There was a positive correlation between the exposure time of all intervention groups and the number of cells in the fibroblast culture. Prolonged exposure time to 0.04% MH, 0.1% MH, and 0.04% TH were positively correlated with the number of keratinocytes. The application of MH during tympanoplasty increased complete TM re-epithelialization through its effect on fibroblasts and keratinocytes proliferation. MH has the potential to improve tympanoplasty outcomes. This present study also illustrated the positive effects of TH on fibroblasts and keratinocytes; thus, its potential therapeutic properties could be further explored.