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"Latar Belakang: Penemuan sel stem jantung (CSC, cardrkzc stem cells) membuktikan jantung sebagai organ dengan pergantian sel-sel parenkim dan non-parenkim di bawah pengaturan kompartemen sel stem. Kemampuan regenerasi jantung berkurang dengan bertambahnya usia. Penyebab penuaan sel stem jantung adalah perubahan pada lingkungan mikro (niche) jantung yang mempengaruhi keberlangsungan hidup sel stem jantung. Tenascin C adalah molekul di niche jantung yang berperan dalam remodeling jaringan jantung dan angiogenesis, dna komponen peuting dalam regenerasi jantung. Penelitian ini bertujuan untuk mengetahui jumlah dan diameter sel otot jantung di jantung tikus yang berbeda usia, menilai ekspresi Tenascin C dan mengetahui hubungan antara ekspresi tenascin C dan perubahan morfometri sel otot jantung. Metode: Desain penelitian ini adalah komparatif potong lintang dengan 6 tikus neonatus (usia 1-4 hari), 9 tikus dewasa muda (usia 3-4 bulan) dan 9 tikus dewasa (usia 12-16 bulan). Proses pembuatan sediaan mikroskopik dilanjutkan dengan pewarnaan HE dan imunohistokimia Tenascin C (sc-9871, sc~2023). Mikrofografi jamung (HE) dipilih 2 Ipb atrium dan 2 Ipb ventrikel. Hasil mikrofotograf dimasukkan dalam format jpeg dan dianalisis dengan Digimizer Image Analyzer. J umlah sel otot jantung dihitung per Ipb dengan tagging system dan diameter sel otot jantung diukur berdzsar unit kalibrasi skala mikrometer. Milcrofotograf tenascin C Iewat software DP2BSW dalam format tifl Dihitung 100 sel otot jantung atrium dan 100 sol ventrikel untuk musing-masing subyek. Imunoreaktivitas tenascin C di sel otot jantung dinyatakan lokasi ekqaresi dan skor intensitas. Lokasi ekspresi adalah positif intra sel, ekstra seL kombinasi keduanya dan negatifl [ntensitas pewarnaan tenascin C diberi skor 1 (lemah) sampai 3 (lcuat). Analisis statistik menggunakan SPSS 13. Hasil : Jumlah sel otot jantung per Ipb terbesar di kelompok neonatus (Atrium=73.4:l=4.8'7; Ventrikel= l52.5:1:3.6) dan paling sedikit di kelompok dewasa (Atrium= 26:I:1.5; Ventrikel= 43.7:1:2.8). Diameter sel otot jantung terkecil di kelompok neonatus (Atrium= 6.lprni0.23; Ventrikel=-° 7.39pmi0.3) dan paling besar di kelompok dewasa (Atrium°-= l7.42pmi0.42; Ventrikel== 23.44|1m=1=0.74). Ekspresi tenascin C ditemukan pada jantung tikus neonatus, dewasa ruuda dan dewasa. Pola ekspresi tenascin C yang sering ditemukan di kelompok neonatus adalah pola kombinasi (Atrium= 43.l7i9.4, Ventrikel= 56.83=l=8.5) dan pola intra sel (Atrium-= 41.33=+=13.4; Ventrikel= 33 .67:|:6.7). Pola ekspresi tenascin C ekstra se! lebih sering ditemukan di kelompok dewasa muda (Atrium= ll.56t3.2; Ventrikel= l2.ll=b7.4) dan dew:-lm (Atrium= 9.22=l:3.5; Ventrikel= 11.67:E3.9) dibandingkan kelompok neonatus (Atrium= 3.33:I=1.3; Ventrikel= 2.5¢l.4). Ekspresi tenascin C negatif paling sering ditemukan di ventrikel jantung dewasa muda (74.44t8 2) dan dewasa (67 .33=\:?7 .6) . Intensitas pewamaan tenascin C kuat (skor 3) paling sering ditemukan di kelempok neonatus (Atrium= 42.83=1=l3.6; Venti-ikel= 59.33=1=9). Skor I paling sering ditemukan di ventrikel jantung kelompok dewasa (16.1 l=|=5.3). Dari analisis korelasi bivariat Pearson ditemukan korelasi positif yang bermakna antara pola ekspresi tenascin C kombinasi di atrium dengan jumlah sel ototjantung atrium (p==0.0l6); pola ekspresi tenasein C intra sel di ventrikel dengan jumlah sel otot jantung ventrikel (p=0.0l) dan pola ekspresi kombinasi di ventrikel dengan jumlah (p=0.00) dan diameter sel otot jantung vcntrikel (p=0.026). Ditemukan pula korelasi positif yang bermakna aniara sl-cor 3 intensitas pewarnaan tenascin C di atrium dengan jumlah sel otot jamung atrium (p=0.035); skor 3 di ventrikel dengan jumlah sei otot jantung ventrikel (p=0.00). Korelasi negatif yang bermakna ditemukan antara skor 3 di ventrikel dengan diameter sel otot jantung ventrikel (p=0.0~0l).Kesimpulan : Semakin bertambah usia jantung, jumlah sel/Ipb semakin berkurang dan diameter semakin besar. Gambaran ini menandalcan teajadinya hipertrofi sel otot jantung. Ekspresi tenascin C ditemukan di jantung neonatus, dewasa muda dan dewasa. Semakin bertambah usia jantung terjadi penurunan jumlah sel otot jamung yang positif mengekspresikan tcnascin C dan berkurangnya intensitas pewarman tenascin C. Di atrium dan ventrikel jamung, semakin banyak jumlah sel otot dengan pola ekspresi tenascin C kombinasi maka semakin banyak jumlah sel otot jantung. Di ventrikel, pola ekspresi kombinasi juga berkorelasi positif dengan diameter se] otot jantung. Semakin tinggi jumlah sel dengan skor intensitas 3 make jumlah sel Otot jantung semakin banyak dan diameter sel otot jantung yang kecil.

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Background: Discovery of Cardiac Stem Cells (CSC) showed the heart as renewable organ with parenchymal and non-parenchymal cells turnover governed by stem cells compartments. Cardiac regenerative ability decreases with advancing age. The cause of CSC?s aging is the changes in cardiac microenvironment (niche) that surrounds CSC. Tenascin C is a major glycoprotein in cardiac niche that plays a vital role in cardiac remodelling and angiogenesis, two main components of cardiac regeneration. This study aims to compare immunoreactivity of tenascin C, cardiomyocites number and diameter in three age groups rat cardiac and determine the correlation between tenascin C immunoreactivity and cardiomyocite?s motphometric changes.

Methods: Design of this study is comparative cross sectional with 6 neonate rats (age I-4 days), 9 young adult rats (age 3-4 months), and 9 adult rats (age 12-16 months). The subjects underwent intravital lixation and cardiac organ was removed. Microscopic specimens were made and stained with hematoxylin-Eosin and tenascin C immunohistochemistry (sc-9871, sc-2023). From cardiac microphotograph (HE stained) two high power field (hpf) was selected for atrium and two hpf for ventricle. Microphotographs was transferred into digital format (jpeg) and analysed with Digimizer Image Analyzer. Cardiomyocite number was determined using tagging system and measurement of cardiomyocite diameter was calibrated with micrometre scale using Digimiaer Image Analyzer. immunohistochemistry results were documented with DPZBSW as tnicrophotographs in digital format (tiff). 100 catrliomyocites in the atrium and in the ventricle fiom each subject was analysed. Immunoreactivity of tenascin C was classified based on expression paltem and staining intensity. The expression pattern was positive intra cellular, positive extra cellular, positive combination (both intra and extra cellular) and negative. Staining intensity was scored I (weak) to 3 (strong). Statistical analysis was performed with SPSS I3.

Result : The most abundant cardiomyocte number per high power fielf (hpf) was found in neonate cardiac (Atrium= 73.4=b4.8'7; Ventrikel= l52.5:l:3.6) and the least abundant was in adult cardiac(Atrium= 26=l:l.5; Ventrikel= 43.7=E2.8). Cardiomyocite diameter was smallest in neonate cardiac (Atrium= 6.1 um=h0.28; Ventrikel= 7.39um:I=0.3) and largest in adult group (Atrium= l7.42um:1:0.42; Ventrikel= 23.44|.un:l:0.74). Tenascin C immunoreactivity was found in neonate, adolescence and adult cardiac. Tenascin C expression pattern most frequently found in neonate cardiac was positive combination (Atrium= 43.l7:1:9.4, Ventrikel= 56.83=l:8.5) and positive intra cellular (Atrium= 4l.33il3.4; Ventrikel= 33.67=l:6.'7). Tenascin C positive extra cellular was commonly found in young adult cardiac (Atrium= 1l.56=l=3.2; Ventrikel= 12.l 1174) and adult cardiac (Atrium= 9.22d:3.5; Ventrikel= 11.671-3.9). Negative tenascin C was more ti-equently found in young adult ventricle (74.44=i=8.2) and adult cardiac (67.33:l:'7.6). High score for tenascin C staining intensity (score 3) was iiequently found in neonate cardiac (Atrium= 42.83=kl3.6; Ventrikel= 59.33d=9). Score l was iiequently found in adult ventricle (16.1l:l:5.3). Pearson bivariate correlation revealed significant correlation between positive combination tenascin C pattern in the atrium with atrial cardiomyocites number(p=0.0l6); positive intra cellular tenascin C pattem in the ventricle with ventricular cardiomyocitcs number (p=0.0l) and positive combination in the ventricle with ventricular cardiomyocites number (p=0.00) and diameter (p=0.026). Significant correlation was also found between score 3 in the atrium with atrial cardiomyocites number (p=0.035); score 3 in the ventricle with ventricular cardiomyocites number (p=0.00). Negative correlation was found signiiicant between score 3 in the ventricle with ventricular cardiomyocitcs diameter (p=0.00l).

Conclusions : With advancing age, cardiomyocte number per hpf decreases while the diameter increases. This resembles hypertrophy of cardiomyocite. Tenascin C immunoreactivity was found in neonate, adolescence and adult mrdiac tissue. With advancing age, we found reduced number of cardiomyocites expressing tenascin C and decreased staining intensity. In cardiac atrium and ventricle, increased number of positive combination tenascin C expression showed increased cardiomyocites number. In ventricle, increased number of positive combination showed increased cardiomyocitec diameter. Increased number of cardiomyocites with score 3 tenascin C staining intensity showed higher cardiomyocites number and smaller diameter."

"Latar Belakang: Sirkumsisi adalah prosedur bedah untuk menghilangkan kulit prepusium. Prosedur ini melibatkan proses penyembuhan luka yang meliputi 3 fase: inflamasi, pembentukan jaringan, dan remodeling jaringan. Tenascin-c adalah protein matriks ekstraselular yang diekspresikan pada saat perlukaan, perbaikan, dan regenerasi jaringan. Tenascin-c ditemukan pada area inflamasi, terutama di tepi perlukaan. Riset ini bertujuan mengidentifikasi korelasi antara ekspresi tenascin-c pada tepi luka sirkumsisi dengan resolusi inflamasi pada penyembukan luka sirkumsisi.Metode: Sampel preputium didapatkan dari kegiatan sirkumsisi masal, kemudian sampel melalui histotechniques dan immunohistokimia spesifik untuk tenascin-c. Data juga diperoleh dari wawancara yang dilaksanakan 14 hari setelah sirkumsisi. Wawancara diikuti oleh observasi fisik untuk menentukan resolusi inflamasi pada perlukaan pasien.Hasil: 85,7 dari sampel yang tenascin-c positif mengalami resolusi inflamasi yang normal. 66,67 dari sampel yang tenascin-c negatif mengalami resolusi inflamasi yang tertunda.Kesimpulan: Ada korelasi antara ekspresi tenascin-c dan resolusi inflamasi pada perlukaan pasca sirkumsisi.

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Background Circumcision is a common invasive surgical procedure to remove the preputial skin. It involves the wound healing process, consisting of 3 phases inflammation, tissue rebuilding, and tissue remodeling. Tenascin c is an extracellular matrix protein highly expressed during tissue injury, renewal, and regeneration. Tenascin c expressions are found at sites of inflammation, it especially peaks at the incision wound edges. This research aims to identify a correlation between tenascin c expressions at the circumcision incision area and the inflammation resolution of circumcision wound healing.Method Preputial skin samples were obtained from a mass circumcision event, afterwards they underwent histotechniques which includes hematoxylin eosin staining and immunohistochemistry specific for tenascin C. Data was also obtained from a follow up interview conducted 14 days after the surgical procedure. The interview was confirmed with physical observation to determine state of inflammation resolution.Results 85,7 of tenascin c positive samples exhibits normal inflammation resolution. 66,67 of tenascin c negative samples exhibit delayed inflammation resolution.Conclusion There is a correlation between tenascin c expression and inflammation resolution in post circumcision wound healing."

"ABSTRACTSirkumsisi telah dianggap sebagai salah satu prosedur bedah tertua dan paling sering dilakukan. Meskipun telah banyak teknik-teknik sirkumsisi yang telah diciptakan, tidak ada mufakat dalam penentuan metode sirkumsisi yang terbaik dalam praktik, terutama dalam ketentuan perbaikan jaringan kulit. Tenascin-C TNC adalah glikoprotein ekstraselular yang terbentuk selama embriogenesis dan meningkat sewaktu penyembuhan luka, terutama dalam fase resolusi. Dalam penelitian ini, penulis menganalisa penyembuhan luka akibat sirkumsisi dengan dorsal-slit method dan kauter laser melalui ekspresi Tenascin-c. Kulit khatan dari 20 partisipan laki-laki 5-12 tahun dikumpulkan dan diwarnai dengan pewarna hematoxylin-eosin untuk menentukan area insisi. Ekspresi tenascin-c diamati dengan imunohistokimia: rasio area dengan TNC positif dan batas pinggiran insisi. Hal ini diikuti dengan daftar pertanyaan pasca-operasi beserta foto-foto dari partisipan untuk menentukan status penyembuhan luka. Grup konvensional memperlihatkan ekspresi TNC yang lebih besar 57.28 47.56 dibanding grup kauterisasi 25.36 16.44 p=0.07 . Rata-rata ekspresi TNC pada subyek dengan penyembuhan luka yang normal 42.15 40.87 sedikit lebih tinggi daripada rata-rata pada subyek dengan penyembuhan luka yang tertunda 38.83 33.40 p=0.872 . Tidak ada korelasi yang signifikan antara ekspresi tenascin-c dengan proses penyembuhan luka. Dari data yang terkumpul dapat dilihat bahwa dorsal-slit dan kauterisasi method, kedua-duanya membuahkan perbaikan jaringan kulit yang normal.

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ABSTRACTCircumcision has been noted as one of the oldest and most common surgical procedure. Even though, various techniques have been developed, there is no consensus on best practice method for circumcision in terms of skin tissue repair. Tenascin c TNC is an extracellular glycoprotein expressed during embryogenesis and markedly increased in wound healing, especially in resolution phase. In this study, the author analyzed the outcome of skin tissue repair dorsal slit and laser cauterization through expression of tenascin c. Prepuces from 20 male participants 5 12 years old were collected and stained using hematoxylin eosin staining to determine incisional area. Tenascin C expression was determined by immunohistochemistry with ratio of TNC positive area and incisional margin. Follow up investigation was done using post operative questionnaire and photographs to determine the status of wound healing. The conventional group showed greater TNC expression 57.28 47.56 than cauterization group 25.36 16.44 p 0.07 . The mean expression of TNC in normal wound healing subjects 42.15 40.87 is slightly more than the mean of delayed wound healing subjects 38.83 33.40 p 0.872 . There is no significant correlation between tenascin c expression and wound healing process. The number of subjects with normal healing after cauterization or conventional techniques is almost identical. The data presented here suggested that both dorsal slit and cauterization methods resulted in normal skin tissue repair. "

"ABSTRAKLatar Belakang: Transplantasi sel sumsum tulang dilaporkan memperbaiki fibrosis hati. Beberapa studi in vitro menunjukkan bukti mekanisme perbaikan dengan melakukan ko-kultur 2D sel sumsum tulang dan sel stelata hepatik. Pada studi tersebut, sel sumsum tulang menghambat aktivasi sel stelata hepatik dan mengurangi deposisi matriks ekstra sel. Pada penelitian ini, mekanisme perbaikan tersebut diteliti dengan melakukan ko-kultur sel sumsum tulang dan sel stelata hepatik pada model kultur 3D dan meneliti efeknya terhadap ekspresi tenascin-C, suatu glikoprotein matriks yang memiliki kontribusi dalam fibrogenesis hati.Metode: Sel stelata hepatik dan sel sumsum tulang yang diisolasi dari tikus dikultur sendiri (monokultur) dan diko-kultur direk dengan metode hanging drop. Karakterisasi sel sumsum tulang dilakukan dengan analisis flowcytometry CD90CD34. Sampel dari kedua kelompok kultur dipanen pada hari ke-7 untuk analisis imunositokimia tenascin-C.Hasil: Persentase sel CD90+CD34- dari sel sumsum tulang yang diisolasi adalah 35,2%. Hasil yang diperoleh pada penelitian ini menunjukkan bahwa sel sumsum tulang memilki efek antifibrotik yang dibuktikan dengan penurunan signifikan ekspresi tenascin-C pada kelompok ko-kultur (p < 0,05) dibandingkan dengan kelompok monokultur pada hari kultur ke-7.Kesimpulan: Temuan tersebut menunjukkan bahwa sel sumsum tulang memiliki efek terapeutik potensial terhadap proses fibrosis hati melalui efeknya dalam meminimalkan ekspresi matriks ekstra sel tenascin-C.

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ABSTRACT

Background: Transplantation of bone marrow derived cells (BMCs) has been reported to improve liver fibrosis. Several in vitro studies have shown evidence for the mechanism of improvement by co-culturing BMCs and hepatic stellate cells (HSCs) in 2D models. In those studies, BMCs were reported to inhibit HSCs activation and reduce extracellular matrix deposition. In this study, we investigated the mechanism by co-culturing BMCs and HSCs in 3D model and its effect on tenascin-C expression, an extracellular matrix glycoprotein that has a contribution in liver fibrogenesis.

Methods: Primary isolated rat HSCs and BMCs were cultured alone (monoculture) and directly co-cultured with hanging drop method. Characterization of BMSCs was performed by flowcytometry CD90CD34 analysis. The monoculture and co-culture samples were harvested on day 7 for tenascin-C immunocytochemistry.

Results: The percentage of CD90+CD34- cells from the isolated BMCs was 35.2%. Result of the present study showed that BMCs have a significant antifibrotic effect as evidenced by the significant decrease in in tenascin-C expression in the co-culture group (p < 0.05) compared to the monoculture group on day 7.

Conclusions: This finding demonstrates that BMSCs have a potential therapeutic effect against liver fibrotic process through their effect in minimizing extracellular matrix tenascin-C expression."

"ABSTRAKPenggunaan sel punca sebagai anti fibrosis hati cukup menjanjikan. Sel punca CD34 asal darah tali pusat sudah banyak digunakan dalam studi anti fibrosis hati. Penelitian ini menjelaskan efek ko-kultur antara sel stelata hepatik HSC LX-2 dan sel punca CD34 asal darah tali pusat dalam morfologi sel dan ekspresi TGF-?, tenascin-C dan kolagen tipe 1A1. Metode : Sel CD34 diisolasi dari sel darah tali pusat manusia yang dikriopreservasi menggunakan separasi magnet. Sel HSC LX-2 dikultur sebagai kontrol monokultur. Sebagian dipanen dan dihitung untuk dilakukan ko-kultur dengan sel CD34 dalam rasio 1:1. Ko-kultur CD34 dan LX-2 dilakukan dengan metode kultur konvensional 2D dan 3D hanging drop. Hasil monokultur dan ko-kultur dipanen pada hari ke1, 2 dan 3 dan dilakukan pewarnaan imunositokimia tenascin-C ekstraksi RNA untuk analisis kuantitatif dengan real time PCR ekspresi TGF-? dan kolagen tipe 1A1.Hasil : Hasil menunjukkan perbedaan morfologi ko-kultur 2D dan 3D hanging drop dibandingkan kontrol monokultur. Pada ko-kultur 2D terdapat mikromassa, sedangkan pada monokultur 2D tidak ada mikromassa yang terbentuk. Pada ko-kultur 3D hanging drop, terdapat spheroid yang lebih kecil hambatan pembentukan spheroid dibandingkan monokultur 3D hanging drop. Sel CD34 memiliki efek direk terhadap aktivitas sinyal sel stelata hepatik dengan adanya kecenderungan penurunan ekspresi TGF-?. Analisis imunositokimia tenascin-C dalam mikromassa dan spheroid masih perlu dioptimasi. Ko-kultur 2D dan 3D hanging drop method sel punca CD34 asal darah tali pusat dan sel stelata hepatik memiliki efek terhadap penurunan ekspresi kolagen tipe 1A1.Kesimpulan : Sel punca CD34 asal darah tali pusat memiliki efek direk terhadap morfologi sel, inhibisi aktivitas sel stelata hepatik LX-2 yang ditandai dengan penurunan ekspresi TGF-beta dan inhibisi deposisi matriks ekstrasel yang ditandai penurunan ekspresi kolagen tipe 1A1.Kata kunci: sel punca asal darah tali pusat CD34 , sel stelata hepatik, liver fibrosis, TGF-beta, tenascin-C, kolagen 1A1.

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ABSTRACTBackground The development of stem cell therapy antifibrotik placing as one of the promising therapy. Umbilical cord blood CD34 stem cells has been widely used in the study antifibrosis. This study describes the effect of co culture between hepatic stellate cells HSC LX 2 and umbilical cord blood CD34 stem cells on cell morphology and expression of TGF , tenascin C and collagen type 1A1.Method CD34 cells were isolated from thawed cryopreserved human umbilical cord blood cells using magnetic separation. LX 2 cells culture were harvested and counted. CD34 and LX 2 cells were mixed in suspension with 1 1 ratio v v . Cell suspension divided into 2 sets 2D co culture plated in standard well plate and 3D co culture as hanging drops. LX 2 monoculture, CD34 dan LX 2 coculture were harvested on day 1, 2 and 3 as sample for further analysis. Tenascin C expression was analysed by imunocytochemistry techniques. TGF Beta and collagen type 1A1 expression was analysed by qPCR.Result The result showed different morphology between co culture and monoculture on 2D and 3D hanging drop. The 2D co culture showed micromass formation, instead of no micromass formation on monoculture. The 3D hanging drop showed smaller spheroid formation spheroid formation inhibition compared with monoculture. CD34 cells showed direct effect on hepatic stellate cell signalling activity represented by the decrease in TGF beta expression, inhibition of extracellular matrix deposition represented by a decrease in Collagen type 1A1 expression.Conclusion UCB CD34 cells showed direct effect on cell morphology, inhibition of hepatic stellate cell LX 2 activity represented by a decrease in TGF beta expression, inhibition of extracellular matrix deposition represented by a decrease in collagen type 1A1 expression. "