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"This book presents the morphological features, dynamics, and sequelae of adenovirus and Thygeson's keratitides captured at high magnification in the living human cornea. It thereby fills the existing void between conventional photographs and slit-lamp observations. Case reports demonstrate the importance of patient history in differential diagnosis, illustrate the need for familiarity with early manifestations of adenovirus infections, and assist in the diagnosis of rare variants of TSPK. Furthermore, the detailed observations on the natural course of the diseases ensure that the book will serve not only as a diagnostic tool but also as a reference when evaluating the effects of potential new treatments."

"It is well known that nifedipine administration in hypertensive patients results in gingival hyperplasia. The aim of this study was to study the pattern of nifedipine-induced gingival hyperplasia, based on morphometric and histological changes as well as on PCNA (Poliferating Cell Nuclear Antigen) expression in the gingival epithelium. In total, 36 male Sprague Dawley rats at the age of 6 - 8 weeks were divided into nine experimental groups and three control groups. Each animal received daily DMSO (dimethyl sulfoxide) via oral intubation at a dosage of 0 (for control groups), 15, 30 or 60 mg/kg (experimental group) of body weight for 7, 21 or 42 days. After the animals were sacrificed, impression of the lower gingival tissue was taken to measure mesio-distal distance, lanio-lingual distance and papilla height. The number of blood vessels and the thickness of gingival epithelium were assessed from hematoxylin and eosin stained sections. Proliferative activity of the epithelial cells was determined by immunohistochemical analysis using PCNA monoclonal antibody. Significant increase in the mesio-distal and labio-lingual distance of the lower gingival tissue was detected morphometrically (p< 0.05). There were more blood vessels in the experimental groups than in the control groups, however there was no specific pattern based on the dosage or duration of nifedipine administration. On the other hand, significant differences were found in the gingival epithelial thickness and proliferative activity between the experimental and the control groups. PCNA-positive cells were observed in basal and suprabasal layers, but nearly none in lamina propria."

"ABSTRAKPendahuluan. Pemahaman dan publikasi mengenai aspek biologi sel osteosarkoma manusia di Indonesia masih terbatas, sehingga dibutuhkan suatu penelitian tentang isolasi, kultur dan karakterisasi sel osteosarkoma manusia secara in vitro dan pada model hewan secara in vivo. Penelitian ini bertujuan untuk mengetahui apakah sel osteosarkoma manusia dapat diisolasi dan dikultur secara in vitro dan secara in vivo pada hewan model tikus Sprague Dawley (SD).Metode Penelitian. Pada tahap pertama dilakukan isolasi dan kultur sel osteosarkoma dari 6 pasien pre-kemoterapi neoadjuvant dan 4 pasien telah mendapat kemoterapi neoadjuvant. Isolasi dan kultur dengan metode eksplant. Karakterisasi sel osteosarkoma dibuktikan dengan pemeriksaan morfologi sel, reverse transcriptase polymerase chain reaction (RT-PCR), immunofluorescence assay (IFA) dan imunositokimia. Tahap kedua menghasilkan model hewan tikus SD dengan inokulasi sel hasil kultur 1.106 sel per ekor ke intramedular femur distal (3 ekor) dan ke intramuskular gastroknemius dan soleus tibia proksimal (3 ekor). Pengukuran Alkali fosfatase serum dilakukan pada minggu ke-0, 4, dan 8, radiologi pada minggu ke-4 dan ke-8 dan histopatologi dilakukan pada minggu ke-8.Temuan Penelitian. Sitologi menunjukkan sel tumor dengan inti yang pleiomorfik, hiperkromatik, letak di tepi dan anak inti nyata. Pemeriksaan RT-PCR menunjukkan ekspresi gen positif terhadap marker STAT3, Nanog, OCT3/4 dan CD 133. Pada pemeriksaan IFA didapatkan hasil positif terhadap antibodi osteokalsin, alkali fosfatase, dan CD 133. Pada pemeriksaan imunositokimia didapatkan hasil positif terhadap antibodi alkali fosfatase dan osteokalsin. Tahap kedua, evaluasi radiologi tidak menunjukkan gambaran destruksi tulang maupun tumbuhnya massa pada soft tissue. Pada histopatologi gambaran jaringan yang normal.Simpulan. Sel osteosarkoma dapat diisolasi dan dikultur dari jaringan tumor pasien osteosarkoma serta menunjukkan karakterisasi sel sesuai gambaran osteosarkoma pada pasien penderita osteosarkoma. Belum dapat dilakukan pembuatan hewan model osteosarkoma dari hewan Tikus Sprague Dawley immunocompetent.

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ABSTRACTIntroduction. Understanding and publications about biological aspect of human osteosarcoma cells in Indonesia are scarce, so study about isolation, culture and characterization of by in vitro or in vivo are needed. This study aimed to understand whether human osteosarcoma cells could be isolated and cultured by in vitro and in vivo for animal model Sprague Dawley (SD) rat.Methods. First stage, isolation and culture of osteosarcoma cell from 6 patients with neoadjuvant prechemotherapy and 4 that already received neoadjuvant chemotherapy. Isolation and culture used explant method. Characterization used morphological examination of cells, reverse transcriptase polymerase chain reaction (RT-PCR), immunofluorescence assay (IFA) and immunocytochemistry. Objective of second stage was producing animal models experiment by inoculation of 1.106 cells intra medullary distal femur (3 animals) and to intramuscular gastrocnemius and soleus proximal tibia (3 animals). Serum alkaline phosphatase was checked at week 0,4 and 8, radiology week 4 and 8, and histopathology at week 8.Results. Cytology showed tumor cell with pleiomorphic, hyperchromatic nucleus on the edge and conspicuous nucleoli. RT-PCR examination was positive for gene expression in STAT3 marker, Nanog, OCT 3/4 and CD 133. IFA was positive for osteocalcin, alkaline phosphatase, and CD 133 antibodies. In immunocytochemical examination there were positive result of alkaline phosphatase and osteocalcin antibody. For second stage, radiology evaluation showed no bone destruction or mass growth in soft tissue. Histopathology showed normal tissue.Conclusions. Osteosarcoma cell could be isolated and cultured from osteosarcoma?s patient and showed cell characterization that corresponded with the picture of osteosarcoma cell in patient with osteosarcoma. Animal model of osteosarcoma in immunocompetent SD rat could not be done."

"Infark miokard menyebabkan kematian kardiomiosit dan remodeling jantung pada situasi patologis. Pascainfark jantung tidak mampu mengatasi kehilangan kardiomiosit meskipun telah dilakukan rekanalisasi atau revaskularisasi. Oleh karena itu, diperlukan metode untuk mengembalikan fungsi jantung. Sel punca dapat memperbaharui diri dan berdiferensiasi menjadi berbagai tipe sel namun kesintasannya pada pasien masih rendah. Untuk meningkatkan retensi dan regenerasi sel punca di miokardium dapat digunakan perancah/scaffold dan sistem ko-kultur, namun belum ada penelitian tentang hal tersebut. Penelitian ini bertujuan mengembangkan terapi infark menggunakan injeksi hidrogel transepikardial dan implantasi di epikardial perancah patch membran amnion yang dideselularisasi menggunakan amniotic epithelial cells (AEC) dengan ko-kultur kardiomiosit. Penelitian ini menggunakan post-test only control group design yang dilakukan di Institut Pertanian Bogor dan Fakultas Kedokteran Universitas Indonesia dari Juli 2021–Oktober 2022. Subjek penelitian adalah 15 babi Sus scrofa domesticus usia 2-3 bulan dibagi tiga kelompok: pAEC, pAEC + kardiomiosit, kontrol positif, dan 1 babi sebagai kontrol negatif. Torakotomi dilakukan untuk membuat model infark dengan ligasi arteri proximal branch to left ventricle (PLV) dilanjutkan implantasi pAEC dengan atau tanpa ko-kultur kardiomiosit pada kelompok terapi, kemudian diobservasi selama 6–8 minggu. Luas infark diukur dengan late gadolinium enhancement MRI; remodeling ventrikel kiri dengan ekokardiografi untuk menilai kontraktilitas, fibrosis dengan IHK, kardiomiogenesis dan regulasi apoptosis dengan RT-PCR, angiogenesis dinilai dengan IHK, dan fraksi ejeksi dinilai dengan ekokardiografi. Luas infark menurun pada kedua kelompok terapi (2,5 [2,00–3,00]% dan 3,60 ± 1,34% vs 9,50 ± 1,91%). Pewarnaan HE dan Masson trichrome menunjukkan berkurangnya proses fibrosis pada kedua kelompok, dikonfirmasi dengan hiperekspresi kolagen1 yang padat dan kaku pada kontrol positif dibandingkan kedua kelompok terapi yang memiliki ekspresi kolagen3 lebih dominan. Ekspresi α-smooth muscle actin pada kedua kelompok tampak tersebar menunjukkan penurunan fibrosis dan kontrol positif menunjukkan peningkatan fibrosis. Peningkatan kardiomiogenesis pada kedua kelompok dikonfirmasi dengan peningkatan ekspresi gen cardiac troponin T, gen myosin heavy chain, gen Nkx.2.5, gen c-Kit, dan penanda otot fungsional α-actinin. Penurunan apoptosis dikonfirmasi dengan penurunan ekspresi gen modulator apoptosis p21 dan ekspresi gen p53 yang berarti diferensiasi sel punca tidak bersifat tumorigenik. Regulasi apoptosis melalui ekspresi kaspase-9 tidak berbeda bermakna. Peningkatan angiogenesis dikonfirmasi dengan peningkatan ekspresi von Willebrand Factor dan ekspresi α-smooth muscle actin yang tersebar. Ekokardiografi menunjukkan perbaikan regional wall motion abnormality lebih banyak pada kelompok terapi daripada kontrol positif dan fraksi ejeksi tidak berbeda bermakna antar kelompok. Disimpulkan kombinasi injeksi hidrogel transepikardial dan implantasi di epikardial perancah patch membran amnion yang dideselularisasi dengan ko-kultur AEC dan kardiomiosit dapat mengurangi luas infark dan remodelling ventrikel kiri, serta meningkatkan angiogenesis pada babi model infark.

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Myocardial infarction induces cardiomyocyte death and remodelling a pathological condition. The post-infarct heart is unable to deal with cardiomyocyte loss despite recanalization or revascularization. Therefore, a procedure is required to restore cardiac function. Stem cells can self-renew and specialize into multiple cell types however the survival of stem cells in patients is still poor. To promote the retention and regeneration of stem cells in the myocardium, scaffolds and co-culture systems may be applicated, although there are no study findings on this issue. This study aimed to develop myocardial infarction therapy using transepicardial hydrogel injection and epicardial decellularized amniotic membrane scaffold patch implantation using amniotic epithelial cells (AEC) with cardiomyocyte co-culture. This study used a post-test-only control group design performed at the IPB University and the Faculty of Medicine, Universitas Indonesia, from July 2021 to October 2022. The study subjects were 15 Sus scrofa domesticus pigs aged 2-3 months placed into three groups: pAEC, pAEC + cardiomyocytes, positive control, and 1 pig as a negative control. Thoracotomy was conducted to create an infarct model with the proximal branch to left ventricle (PLV) artery occlusion followed by pAEC implantation with or without cardiomyocyte co-culture in the therapy group, then evaluated for 6–8 weeks. Infarct size was determined by late gadolinium enhancement MRI, left ventricular remodeling by echocardiography to evaluate contractility, fibrosis by IHC, cardiomyogenesis and regulation of apoptosis by RT-PCR, angiogenesis was assessed by IHC, and ejection fraction by echocardiography. Infarct size reduced in both therapy groups (2,5 [2,00–3,00]% and 3,60 ± 1,34% vs 9,50 ± 1,91%). HE and Masson trichrome staining demonstrated decreased fibrosis in both groups, confirmed by hyperexpression of dense and stiff collagen 1 in the positive control compared to the two therapy groups with more dominant collagen 3 expressions. The α-smooth muscle actin expression in both groups seemed to be scattered suggesting reduced fibrosis while the positive control showed increased fibrosis. Increased cardiomyogenesis in both groups was confirmed by increased expression of the cardiac troponin T gene, the myosin heavy chain gene, the Nkx.2.5 gene, the c-Kit gene, and the functional muscle marker α-actinin. The reduction in apoptosis has been confirmed by lower expression of the p21 apoptosis modulator gene and p53 gene expression, which suggests that stem cell differentiation is not tumorigenic. The control of apoptosis by caspase-9 expression was not significantly different. Increased angiogenesis was verified by increased von Willebrand Factor expression and scattered expression of α-smooth muscle actin. Echocardiography showed greater improvement in regional wall motion abnormalities in the therapy groups than in the positive control, and the ejection fraction was not significantly different between groups. It was concluded that the combination of transepicardial hydrogel injection and epicardial decellularized amniotic membrane scaffold patch implantation using AEC with cardiomyocyte co-culture could reduce infarct size and left ventricular remodeling, as well as increase angiogenesis in infarct model pigs."

"The aim of the study was to compare ex vivo the toxic effects of six root canal sealers immediately after mixing or setting on human periodontal ligament fibroblasts (HPdLF). Freshly mixed (I group) or set (allowed to dry for 24 h) (II group) specimens of AH Plus Jet (AH), Apexit Plus (AP), MTA Fillapex (FL), GuttaFlow (GF), MetaSEAL Soft (META), and Tubli-Seal (TS) were prepared. HPdLF were exposed for 24 h to the specimens. 3-(4,5-dimethylthiazolo-2-yl)-2,5-diphenyltetrazolium bromide assay was used to examine the effect of the root canal sealers on mitochondrial metabolic activity. Fluorescein isothiocyanate (FITC)-annexin V (AnV) and propidium iodide staining followed by flow cytometry was used to identify the effects of the materials on cell apoptosis/necrosis. Statistical analyses were performed by one-way ANOVA followed by post hoc tests, and significance was determined at P < 0.05. Most materials from the two groups reduced the viability of the cultured cells compared with the control group (P < 0.05). Statistical analysis showed significant differences in HPdLF viability between the individual materials in each group (P < 0.001). AH and AP induced a significant increase in the percentage of apoptotic cells, while TS, FL, and META elevated the proportion of necrotic cells compared with other materials and the controls (p < 0.05). The cytotoxic effects of the tested root canal sealers (both fresh and set) on HPdLF varied. Both forms of sealers were able to cause toxic effects by inducing apoptosis and necrosis in HPdLF. The cytotoxicity of FL, META, TS was mainly associated with necrosis, while AH and AP with apoptosis."

"Lensa manusia berfungsi memfokuskan cahaya teriihat, menyerap UV A dan UV B, sehingga sangat rentan terhadap efek fototoksik cahaya yang diterimanya. Sel epitel adalah bagian Iensa yang memiliki peran penting untuk pertumbuhan, diferensiasi dan homeostasis seluruh Iensa. Sel ini selalu terpapar cahaya sehingga sangat mungkin terganggu oleh radiasi UV yang bersifat mutagenik. Mitokondria adalah penghasil energi dan berperan pada kematian sel serta penuaan. MtDNA sangat rentan terhadap paparan radikal bebas karena tidak memiliki histon pelindung dan kemampuan reparasi yang sangat terbatas, oleh karena itu Iaju mutasi mtDNA Iebih tinggi dibandingkan DNA inti. Pada berbagai jaringan yang menua. ditemukan akumulasi mutasi mtDNA terdelesi. Delesi ini mengakibatkan hilangnya gen mtDNA yang menyandi subunit kompleks respirasi mitokondria (kompleks I, III, IV dan V) serta tRNA dan rRNA mitokondria, sehingga teljadi penurunan fungsi OXPHOS. Deiesi mtDNA ini awalnya dilaporkan terdeteksi pada jaringan otot lurik dan otot jantung, tetapi kemudian diiaporkan pula pada berbagai jaringan lain yang menua. Pertanyaan yang sangat penting dalam ilmu oftalmologi adalah, apakah proses yang sama juga berperan di Iensa mata. MtDNA sel epitel Iensa sangat mungkin mengakumulasi mutasi mtDNA karena sel ini selalu terpapar oleh UV, masa hidupnya cukup panjang dan tidak pemah gugur atau hilang. Sampai sekarang pertanyaan tersebut belum terjawab. Paparan UV pada kuitur Iensa menyebabkan apoptosis sel epitel dan kekeruhan Iensa. Pada Iensa manusia, apoptosis ditemukan pada katarak polaris anterior maupun katarak senilis, sedangkan pada iensa jernih hampir tidak ditemukan adanya apoptosis. Tingkat apoptosis sel epitel lensa dan perubahan fungsi respirasi mitokondria mungkin pula berperan pada penuaan dan proses pembentukan katarak. Hubungan antara penuaan, apoptosis dan pembentukan katarak masih merupakan pertanyaan yang belum teljiawab secara menyeiuruh. Oleh karena itu, tujuan penelitian ini ingin mengkaji apakah perubahan fungsi respirasi mitokondria dan tingkat apoptosis berperan dalam fenomena penuaan sei epitel lensa dan proses kataraktogenesis."

"Latar belakang: Terapi sel punca dikembangkan sebagai alternatif terapi gagal jantung akibat infark miokardium. Bermacam tipe sel dengan berbagai metode implantasi telah banyak dikembangkan tetapi belum mendapatkan hasil optimal. Sel h-AECs (Human Amnion Epithelial Stem Cells) memiliki sifat yang sangat mendukung sebagai sumber sel bagi terapi sel punca pada jantung. Teknologi rekayasa jaringan dengan melakukan ko-kultur kardiomiosit dan h-AECs pada biomaterial scaffold diyakini dapat menjawab permasalahan pada pengembangan terapi sel punca pada gagal jantung.Metode: Penelitian ini adalah studi eksperimental in-vitro dengan penyemaian ko-kultur sel kardiomiosit dan h-AECs ke dalam scaffold patch. Kardiomiosit berasal dari otot ventrikel kanan pasien penderita penyakit tetralogy of Fallots yang dilakukan operasi koreksi TOF. Sedangkan sel h-AECs didapat dari epitel amnion yang merupakan limbah operasi seksio sesarea. Setelah dilakukan karakterisasi pada kardiomiosit dan h-AECs, dilakukan ko-kultur pada scaffold amnion dengan perbandingan densitas penyemaian 1:5 dan 1:6. Evaluasi hasil ko-kultur dilakukan dengan penilaian viabilitas sel, ekspresi gen spesifik kardiomiosit dan uji toksisitas patch.Hasil: Hasil ko-kultur kardiomiosit dan h-AECs tidak terdapat perbedaan bermakna pada rerata jumlah sel viabel pada hari kedua dan kelima (p>0,05). Sedangkan pada hari kedelapan terdapat perbedaan bermakna pada jumlah sel viabel, rasio 1:5 menghasilkan jumlah sel viabel lebih baik dibanding rasio 1:6 (p=0,011). Ekspresi gen spesifik kardiomiosit konsisten tampak pada kelompok rasio 1:6 dan mulai menunjukkan signifikan pada hari kedelapan, terdapat perbedaan bermakna pada ekspresi gen di hari kedelapan, kelompok rasio 1:6 mengekspresikan gen cTnT dan ACTN2 lebih baik dibanding kelompok 1:5 (p=0,000 dan p=0,001). Pada uji toksisitas, tidak terdapat perbedaan bermakna pada jumlah ATP dan kadar TNFα antara kelompok 1:5 dan 1:6.Simpulan: Teknik ko-kultur yang dikembangkan dapat menghasilkan sel kardiomiosit baru. Kelompok rasio 1:6 menghasilkan sel yang memiliki sifat spesifik kardiomiosit lebih baik dibanding kelompok rasio 1:5 tetapi menghasilkan jumlah sel viabel lebih sedikit. Patch hasil ko-kultur tidak bersifat toksik.

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Background: Stem cell therapy was developed as an alternative therapy for heart failure due to myocardial infarction. Various types of cells with various methods of implantation have been developed but have not yet obtained optimal results. h-AECs (Human Amnion Epithelial Stem Cells) have very supportive properties as a source of cells for stem cell therapy in the heart. Tissue engineering technology by co-culturing cardiomyocytes and h-AECs on scaffold biomaterials is believed to be able to answer problems in the development of stem cell therapy in heart failure.

Method: This study is an in-vitro experimental study by seeding co-cultures of cardiomyocytes and h-AECs into the scaffold patch. Cardiomyocytes were derived from the right ventricular muscle of patients with tetralogy of Fallot disease who underwent TOF correction surgery. Meanwhile, the h-AECs cells were obtained from the amniotic epithelium which is the waste from cesarean section. After characterization of cardiomyocytes and h-AECs, co-culture was performed on amnion scaffold with seeding density ratio 1:5 and 1:6. Evaluation of co-culture results was carried out by assessing cells viability, expression of specific cardiomyocytes gen and patch toxicity tests.

Result: The results of co-culture of cardiomyocytes and h-AECs showed no significant difference in the mean number of viable cells on the second and fifth days (p>0.05). While on the eighth day there was a significant difference in the number of viable cells, a ratio of 1:5 resulted in a better number of viable cells than a ratio of 1:6 (p=0.011). Cardiomyocyte-specific gene expression was consistently seen in the 1:6 ratio group and began to show significantly on the eighth day, there was a significant difference in gene expression on the eighth day, the 1:6 ratio group expressed cTnT and ACTN2 genes better than the 1:5 group (p= 0.000 and p=0.001). In the toxicity test, there was no significant difference in the amount of ATP and TNFα levels between the 1:5 and 1:6 groups.

Conclusion: The developed co-culture technique can generate new cardiomyocytes. The 1:6 ratio group produced cells that had better cardiomyocyte-specific properties than the 1:5 ratio group but produced fewer cells. Co-culture of h-AECs and cardiomyocytes on patch was not toxic."