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Abstrak :
Latar Belakang. Sumsung tulang merupakan sumber sel punca mesenkimal SPM yang paling banyak digunakan selain jaringan lemak sebagai sumber pengganti yang menjanjikan. Peningkatan penggunaan SPM membutuhkan kemampuan untuk melakukan subkultur pasase SPM. Untuk mengumpulkan dan menyimpan SPM dalam waktu tertentu tanpa mengubah karakter SPM maka dilakukan kriopreservasi. Penelitian ini bertujuan meningkatkan pemahaman efek pasase terhadap penuaan sel punca mesenkimal sumsum tulang dan jaringan lemak yang dikriopreservasi.Metode. Penelitian ini merupakan studi analitik observasional yang dilaksanakan di UPT-TK Sel Punca RSCM FKUI April 2016 - September 2016. Sampel penelitian adalah sel punca mesenkimal sumsum tulang dan jaringan lemak pasase pertama yang dikriopreservasi 1 dan 2 kali. Dilakukan pengukuran terhadap ukuran sel, viabilitas sel, population doubling time PDT, colony forming unit dan penghitungan persentase sel yang menua. Data pasase dianalisis dengan multiple comparison ANOVA dengan Tukey HSD correction dan student t-test menggunakan program SPSS 23. Hasil. Terdapat perbedaan yang signifikan antara kedua kelompok kriopreservasi SPM sumsum tulang dalam PDT, viabilitas, dan ukuran sel pada P6 dengan p ...... Introduction. Bone marrow is still the gold standard source of MSC, but adipose tissue became a promising alternative source. Passage and cryopreservation are effective ways to multiply, pool and store MSC without altering its function. The aim of this research was to enhance the knowledge of the effect of passage on senescence profile of cryopreserved human bone marrow and adipose derived MSC.Method. This research was an observational analytic study to analyze population doubling time PDT, cell size, viability, colony forming unit and percentage of senescent cells and done in UPT ndash TK Sel Punca RSCM FKUI, during April to September 2016. The samples were bone marrow and adipose MSC at passage one, which were cryopreserved for the first and second time. Cryopreservastion groups were analyzed using student t test while inter passage was analyzed using ANOVA test. Result. There were significant differences between both cryopreserved bone marrow groups in PDT, viability and cell size in P6, p

Abstrak :
[ABSTRAK
Pendahuluan. Sel punca mesenkimal (SPM) sangat menjanjikan dalam bidang rekayasa jaringan karena sifatnya yang multipoten, cepat berproliferasi, dan berkemampuan tinggi untuk beregenerasi. SPM sumsum tulang dapat menjadi terapi pilihan nekrosis avaskular (AVN) kaput femur yang banyak diderita oleh pasien lupus eritematosus sistemik (LES) pada masa sekarang ini. SPM sumsum tulang penderita LES mengalami gangguan fenotip, proliferasi, diferensiasi. Terapi SPM pada AVN kaput femur dapat menggunakan donor otologus yang dilaporkan memberikan hasil luaran yang baik dan keamanan yang signifikan. Oleh karena itu, diperlukan penelitian untuk mengetahui potensi, karakteristik, dan diferensiasi SPM sumsum tulang pasien LES yang dihubungkan dengan usia. Metode. Penelitian ini adalah penelitian in vitro yang meneliti 4 subjek penderita LES di Rumah Sakit Cipto Mangunkusumo Jakarta. Aspirat SPM sumsum tulang dilakukan isolasi, ekspansi dan diferensiasi. Analisis statistik menggunakan uji korelasi spearman untuk melihat hubungan usia pasien LES dengan waktu konfluensi, jumlah sel konfluens dan waktu diferensiasi osteogenik, kondrogenik, dan adipogenik. Hasil dan Diskusi. Rerata jumlah sel konfluens adalah 7.44 x 105 ± 3.06 x 105 sel/ml, rerata waktu konfluens adalah 20.75 ± 4.99 hari, median waktu diferensiasi adipogenik yaitu 17.5 hari (rentang 14-21), waktu diferensiasi osteogenik dan kondrogenik yaitu 21 hari. Terdapat korelasi positif bermakna antara usia penderita LES dengan waktu konfluens SPM (p<0.001) dan korelasi negatif bermakna antara usia penderita LES dengan jumlah sel konfluens SPM (p<0.001). Simpulan. SPM sumsum tulang krista iliaka penderita LES mampu diisolasi, berproliferasi dan berdiferensiasi. SPM sumsum tulang penderita LES memiliki waktu konfluens dan waktu diferensiasi yang lebih lama dan jumlah sel konfluens yang lebih sedikit.

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ABSTRACT
Introduction. Mesenchymal stem cells (MSC) is very promising in the field of tissue engineering because it is multipotent, rapidly proliferate, and high ability to regenerate bone marrow. BM-MSC may be treatment of choice of avascular necrosis (AVN) of femoral head that affects many systemic lupus erythematosus (SLE) patients at the present time. BM-MSC of SLE patients has impairment in phenotype, proliferation, and differentiation. Mesenchymal stem cell therapy on femoral head AVN which use autologous donors are reported deliver good outcomes and safety. Therefore, research is needed to determine the potency, characteristics, and differentiation of BM-MSC in patients with SLE and related with age. Methods. This study is in vitro study that examined four subjects as SLE patients in Cipto Mangunkusumo Hospital. BM-MSC of SLE patients is performed isolation, expansion and differentiation. Statistical analysis using pearson and spearman correlation test to see the correlation of age of SLE patients with confluence time, the number of confluence cells and differentiation time. Result and Discussion. Mean of confluent cell numbers is 7.44 x 105 ± 3.06 x 105cells/ml, mean of confluent time is 20.75 ± 4.99 days, median of adipogenic differentiation time is 17.5 days (range 14-21), osteogenic and chondrogenic differentiation time is 21 days. There is a positive correlation between patient?s age with confluence time (p <0.001) and negative correlation with MSC confluence cell count (p <0.001). Conclusion. BM-MSC form iliac crest in patients with SLE can be isolated, proliferated and differentiated. BM-MSC of SLE patients has longer confluence time and differentiation time and lower confluence cell count., Introduction. Mesenchymal stem cells (MSC) is very promising in the field of tissue engineering because it is multipotent, rapidly proliferate, and high ability to regenerate bone marrow. BM-MSC may be treatment of choice of avascular necrosis (AVN) of femoral head that affects many systemic lupus erythematosus (SLE) patients at the present time. BM-MSC of SLE patients has impairment in phenotype, proliferation, and differentiation. Mesenchymal stem cell therapy on femoral head AVN which use autologous donors are reported deliver good outcomes and safety. Therefore, research is needed to determine the potency, characteristics, and differentiation of BM-MSC in patients with SLE and related with age. Methods. This study is in vitro study that examined four subjects as SLE patients in Cipto Mangunkusumo Hospital. BM-MSC of SLE patients is performed isolation, expansion and differentiation. Statistical analysis using pearson and spearman correlation test to see the correlation of age of SLE patients with confluence time, the number of confluence cells and differentiation time. Result and Discussion. Mean of confluent cell numbers is 7.44 x 105 ± 3.06 x 105cells/ml, mean of confluent time is 20.75 ± 4.99 days, median of adipogenic differentiation time is 17.5 days (range 14-21), osteogenic and chondrogenic differentiation time is 21 days. There is a positive correlation between patient’s age with confluence time (p <0.001) and negative correlation with MSC confluence cell count (p <0.001). Conclusion. BM-MSC form iliac crest in patients with SLE can be isolated, proliferated and differentiated. BM-MSC of SLE patients has longer confluence time and differentiation time and lower confluence cell count.]

Abstrak :
Pendahuluan: Efek hiperglikemik dan produk Advanced Glycation Endproduct (AGE) dari diabetes mellitus (DM) sering dikaitkan dengan komplikasi muskuloskeletal seperti neuropati perifer dan tendinopati Achilles pada regio pergelangan kaki. Hal ini beresiko menimbulkan efek lanjutan berupa perubahan struktur berjalan, kekakuan sendi hingga luka tukak telapak kaki. Tatalaksana tendinopati DM hingga saat ini terbatas pada pengurangan gejala lanjutan tanpa meningkatkan proses regenerasi tendon, sehingga dibutuhkan penelitian untuk menilai efek terapi dari sekretom dan eksosom SPM dalam hal perbaikan struktur tendon. Hal ini diwakili oleh penggunaan hewan coba tikus SD yang telah terinduksi menjadi tendinopati DM. Metode: Studi ini melibatkan fase studi pilot pertama, kedua, dan penelitian utama. Tikus SD diperoleh dan diberikan diet tinggi lemak (HFD) dan pemberian larutan fruktosa 55% selama delapan minggu. Diabetes diinduksi menggunakan injeksi streptozotocin (STZ) intraperitoneal berbagai dosis. Studi pilot pertama bertujuan untuk menentukan volume cairan yang dapat diinjeksikan ke area peritendon. Sementara itu, studi pilot kedua bertujuan untuk mengidentifikasi dosis STZ yang efektif. Dalam fase penelitian utama, tikus diabetes menerima injeksi lokal eksosom, sekretom, atau kombinasinya. Setelah perawatan, tikus dieutanasia, dan tendon Achilles dianalisis secara histopatologi dan imunohistokimia. Hasil dan Diskusi: Studi pilot pertama menyimpulkan bahwa 0,8 ml merupakan volume cairan optimal yang dapat diinjeksikan ke area peritendon. Sementara itu, studi pilot kedua menunjukkan bahwa setelah 8 minggu HFD, pemberian fruktosa, dan injeksi STZ, kelompok STZ 26 mg/kg memiliki kadar glukosa 220,54 ± 9,11 mg/dL, dan kelompok STZ 30mg/kg memiliki 213,88 ± 8,99 mg/dL dengan perbedaan paling signifikan dalam skor Bonar diamati di kelompok STZ 30mg/kg, hal ini menunjukkan keberhasilan induksi hewan coba. Pada penelitian utama setelah pemberian sekretom, eksosom, atau kombinasi, kadar TGF-β dan IL-6 dan skor Bonar tidak menunjukkan perbedaan signifikan antar kelompok. Analisis pasca intervensi mengungkapkan perbedaan signifikan dalam kadar IL-6 dan Col-1, dimana pada kelompok perlakuan terdapat penurunan IL-6 yang signifikan pada hari ke-14 dan peningkatan Col-1 yang signifikan pada hari ke-21 dibandingkan dengan kelompok kontrol. Kesimpulan: Penelitian ini menunjukkan bahwa kombinasi diet HFD, pemberian fruktosa, dan dosis injeksi STZ 30 mg/kg efektif menciptakan hewan model tendinopati DM. Skor Bonar yang tinggi pada kelompok STZ mengindikasikan kerusakan tendon signifikan. TGF-β dan IL-6 tidak menunjukkan perbedaan signifikan antar kelompok, namun IL-6 meningkat pada hari ke-14 dan Col-1 pada hari ke-21 pada kelompok intervensi secara signifikan, menunjukkan potensi terapi eksosom dan sekretom pada penyembuhan tendon. ......Introduction: The hyperglycemic effects and Advanced Glycation Endproduct (AGE) of diabetes mellitus (DM) are often associated with musculoskeletal complications such as peripheral neuropathy and Achilles tendinopathy in the region of the legs and ankles. It is one of the risks of developing advanced negative effects such as changes in walking structure, stiffness of the joints to ulcer wounds on the the ankle. The management of DM tendinopathy to date is limited to reducing advanced symptoms without enhancing tendon regeneration process, therefore, further research is needed to assess the therapeutic effects of MSC secretomes and exosomes in terms of tendon structure improvement. It is represented by the use of SD rats induced into DM tendinopathy. Methods: This study involves two pilot study phases and the main research. SD mice were obtained and given a high-fat diet (HFD) and given 55% fructose solution foreight weeks. Diabetes is induced by injection of streptozotocin (STZ). The first phase of the pilot study aims to determine the volume of liquid injected into the peritendon area, and the second phase aims to identify an effective dose of STZ to induce DM. In the main study, diabetic mice received local injections of exosomes, secretomes, or a combination of them. After treatment, the rats were euthanazied, and the Achilles tendon was analysed histopathologically and immunohistochemically. Results and Discussion: The first pilot study concluded that 0.8 ml was the optimal fluid volume that could be injected into the peritendon area. Meanwhile, the second pilot study showed that after 8 weeks of HFD, fructose administration, and injection of STZ, the STZ 26 mg/kg group had a glucose level of 220.54 ± 9.11 mg/dL, and the STZ 30 mg/kg group had 213.88 ± 8.99 mg/dL with the most significant difference in Bonar score was observed in the STZ 30mg/kg group, this indicates successful induction of experimental animals. In the main study after administering secretome, exosome, or a combination of the two, the levels of TGF-β and IL-6 and the Bonar score did not show significant differences between groups. Post-intervention analysis revealed significant differences in IL-6 and Col-1 levels, in which the treatment group there was a significant decrease in IL-6 on day 14 and a significant increase in Col-1 on day 21 compared to the control group. Conclusion: This study shows that a combination of HFD, fructose administration, and STZ 30mg/kg are effective in creating animal model for diabetic Achilles tendinopathy. A high Bonar score in the STZ group indicates significant tendon damage. TGF-β and IL-6 did not show significant differences between the groups, but IL-6 increased on day 14 and Col-1 on day 21 in the intervention groups significantly, indicating the potential for exosome and secretome therapy on tendon healing. Keyword: diabetic Achilles tendinopathy, Sprague Dawley rats, exosome and secretome combination, bone marrow mesenchymal stem cel

Abstrak :
Tendinopati Achilles diabetes merupakan penyakit degeneratif akibat perubahan homeostasis jaringan tendon yang disebabkan oleh diabetes melitus tipe 2. Penyembuhan tendinopati Achilles diabetes sulit untuk dicapai karena terbatasnya kapasitas regenerasi tendon. Eksosom asal sel punca mesenkimal (SPM) sumsum tulang memiliki kemampuan dalam menghambat degenerasi jaringan sehingga berpotensi untuk mengatasi tendinopati Achilles diabetes. Efek eksosom SPM sumsum tulang terhadap tendon Achilles dapat diinvestigasi melalui perubahan ekspresi relatif gen a disintegrin and metalloproteinase domain 12 (ADAM12). Gen ADAM12 merupakan gen pendegradasi matriks yang terekspresi tinggi pada tendinopati Achilles diabetes. Penelitian ini bertujuan untuk mengetahui pengaruh injeksi 0,8 mL eksosom asal SPM sumsum tulang pada tendinopati Achilles tikus diabetes berdasarkan analisis histologi dan ekspresi gen ADAM12. Sebanyak 12 ekor tikus putih jantan galur Sprague Dawley dikelompokkan menjadi dua kelompok yang terdiri atas kelompok kontrol tendinopati (KK) dan kelompok eksosom (KE). Analisis histologi tendon Achilles posmortem hari ke-21 dilakukan dengan metode semikuantitatif skor Bonar dan histomorfometri kuantitatif luas area kolagen melalui pulasan Hematoksilin-Eosin, Alcian Blue, dan Masson’s Trichrome. Perubahan ekspresi gen ADAM12 diperiksa secara kuantitatif menggunakan qRT-PCR. Berdasarkan hasil penelitian, rata-rata skor Bonar KE (1,67 ± 1,282) ditemukan lebih rendah daripada KK (6,40 ± 2,195) secara signifikan (P = 0,001; P < 0,05). Analisis histomorfometri juga menunjukkan rata-rata luas area kolagen KE (85,15 ± 7,023) yang cenderung lebih tinggi dibandingkan KK (76,64 ± 9,237), tetapi tidak berbeda nyata (P = 0,103; P ≥ 0,05). Ekspresi gen ADAM12 KE mengalami perubahan sebesar 0,9 kali lipat lebih tinggi daripada KK, meskipun secara statistik tidak signifikan (P = 0,421; P ≥ 0,05). Dengan demikian, dapat disimpulkan bahwa injeksi 0,8 mL eksosom asal SPM sumsum tulang terbukti memiliki potensi dalam memicu perbaikan tendinopati Achilles diabetes pada hari ke-21. ......Diabetic Achilles tendinopathy is a degenerative disease resulting from changes in tendon tissue homeostasis caused by type 2 diabetes mellitus. The cure of diabetic Achilles tendinopathy is difficult to achieve due to the limited regeneration capacity of the tendon. Exosomes from bone marrow-derived mesenchymal stem cells (MSC) can inhibit tissue degeneration so they have the potential to treat diabetic Achilles tendinopathy. The effect of exosomes from bone marrow-derived MSC on the Achilles tendon can be investigated through changes in the relative expression of a disintegrin and metalloproteinase domain 12 (ADAM12) gene. The ADAM12 gene is a matrix-degrading gene that is highly expressed in diabetic Achilles tendinopathy. This study aims to determine the effect of injection of 0.8 mL of exosomes from bone marrow-derived MSC on Achilles tendinopathy in diabetic rats based on histology analysis and ADAM12 gene expression. A total of 12 male white Sprague Dawley rats were grouped into two groups consisting of the tendinopathy control group (KK) and the exosome group (KE). Postmortem Achilles tendon histology analysis on day 21 was carried out using the semiquantitative Bonar score method and quantitative histomorphometry of collagen area using Hematoxylin-Eosin, Alcian Blue, and Masson's Trichrome staining. Changes in ADAM12 gene expression were examined quantitatively using qRT-PCR. Based on the research results, the mean score of Bonar KE (1.67 ± 1.282) was found to be significantly lower than KK (6.40 ± 2.195) (P = 0.001; P < 0.05). The histomorphometric analysis also showed that the average collagen area of KE (85.15 ± 7.023) tended to be higher than KK (76.64 ± 9.237) but was not significantly different (P = 0.103; P ≥ 0.05). ADAM12 KE gene expression changed 0.9-fold higher than KK, although it was not statistically significant (P = 0.421; P ≥ 0.05). Thus, the injection of 0.8 mL of exosomes from bone marrow-derived MSC was proven to have the potential to trigger improvement in diabetic Achilles tendinopathy on day 21.