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"ABSTRAKPendahuluan. Sel punca mesenkimal merupakan salah satu alternatif pengobatan yang menjanjikan, termasuk dibidang orthopedi. Sumsum tulang masih menjadi pilihan utama sumber sel punca mesenkimal, namun dikarenakan jumlah sel punca mesenkimal yang sedikit, prosedur pengambilan yang invasif dan nyeri, jaringan adiposa mulai digunakan sebagai alternatif dengan kemampuan yang sebanding. Tindakan minimal invasive pada implantasi sel punca pada kasus tulang belakang membutuhkan alat bantu image intensifier C-arm yang menyebabkan sel punca teradiasi sinar X. Penelitian ini bertujuan mengetahui efek pajanan sinar-x c-arm terhadap viabilitas dan potensi osteogenik sel punca mesenkimal dan membandingkan antar kelompok donor. Bahan dan Metode. Penelitian ini merupakan penelitian eksperimental yang dilaksanakan di UPT-TK Sel Punca RSCM januari 2016-februari 2017 . Sampel penelitian adalah sel punca mesenkimal jaringan adiposa dan sumsum tulang pasca kriopreservasi. Sel punca pasca thawing dan propagasi dilakukan pajanan sinar X C-arm dengan berbagai dosis yang dilakukan di Instalasi Bedah Pusat RSUPN Ciptomangunkusumo. Sel punca lalu dikultur dan dilakukan diffenrensiasi osteogenik. Peneliti melakukan analisis viabilitas, waktu penggandaan populasi dan potensi osteogenik dengan pewarnaan alizarin red. Seluruh data dianalisis dengan SPSS 20. Hasil. Tidak terdapat perbedaan viabilitas sel punca mesenkimal jaringan adiposa dan sumsum tulang pre radiasi, pasca radiasi serta pasca radiasi dan kultur pada dosis radiasi yang sama p>0,05 . Tidak terdapat perbedaan potensi osteogenik yang bermakna antara sel punca mesenkimal jaringan adiposa dan sumsum tulang p>0,05 . Terdapat penurunan waktu penggandaan populasi sel punca mesenkimal jaringan adiposa pada dosis radiasi > 5,94 mSv. Kesimpulan. Viabilitas dan potensi osteogenik sel punca mesenkimal sumsum tulang dan jaringan adiposa tidak dipengaruhi oleh paparan sinar X hingga 15,30 mSv. Sel punca mesenkimal jaringan adiposa menunjukkan waktu penggandaan populasi yang lebih pendek pada dosis yang lebih besar. Sel punca mesenkimal jaringan adiposa dan sel punca mesenkimal sumsum tulang memiliki potensi osteogenik yang sebanding

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ABSTRACTIntroduction. Mesencymal stem cells MSCs is a promising alternative treatment in medicine, including in orthopedic. Bone marrow is still the main source for MSCs. Because of relative less stem cell number, limited source, pain and invasive procedure to obtain the bone marrow, adipose tissue is also considered as a valuable source of MSCs with equal potency. Minimally invasive MSC injections in spine need image intensifier C arm as guidance that potentially influence the cell viability and osteogenic potency. The aim of this study is to evaluate the radiation effects from C arm on the viability and osteogenicity among two types of MSCs. Material and Methods. This experimental study was held on Stem Cell Medical Technology Integrated Service Unit Cipto Mangunkusumo Hospital January 2016 February 2017 . Study samples were Adipose Tissue derived MSCs AT MSCs and Bone Marrow MSCs BM MSCs , which had undergone cryopreservation. After thawing and propagation process, we gave x ray radiation with a variety of doses to MSCs at the Operation Theater Cipto Mangunkusumo Hospital. After the radiation, MSCs was took back to the laboratory for culture and osteogenic differentiation. Author analyzed the viability, population doubling time, and osteogenic potential by alizarin red stain. All data were analyzed using SPSS 20. Results. There was no significant difference among MSCs groups in term of cell viability before radiation, after radiation, and after radiation and culture p 0.05 . There was also no significant difference of the osteogenic potential between the two MSCs groups p 0.05 . However, there was a reduction in population doubling time of AT MSCs radiated with more than 5.94mSv radiation dose. Conclusions. Viability and osteogenic potential of either AT MSCs or BM MSCs were not affected by x ray radiation up to 15.3 mSV. AT MSCs showed a shorter population doubling time when given larger radiation dose. AT MSCs and BM MSCs had equal osteogenic potency. "

"[ABSTRAKPendahuluan: Sel punca mesenkimal (SPM) sebagai salah satu alternatif terapi kasus sulit dapat diperoleh dari jaringan adiposa. Kemampuan SPM dalam rekayasa jaringan membutuhkan prosedur implantasi SPM yang aman dan bebas kontaminasi. Tindakan minimal invasive pada kasus cedera medulla spinalis dengan terapi implantasi SPM dapat menyebabkan sel tersebut terpajan radiasi sinar-x c-arm. Viabilitas dan waktu penggandaan populasi (WPP) merupakan salah satu komponen utama keberhasilan prosedur implantasi tersebut. Penelitian ini bertujuan mengetahui efek pajanan sinar-x c-arm terhadap viabilitas dan WPP SPM.Metode: Penelitian ini adalah penelitian eksperimental SPM jaringan adiposa pasca cryopreservation. Sel punca pasca thaw dan propagasi kemudian dilakukan pajanan radiasi sinar-x dengan c-arm. Sel punca kemudian di kultur untuk menilai viabilitas dan waktu penggandaan populasi. Uji Generalized Linear Model untuk menilai perbedaan viabilitas antara besar dosis radiasi. Uji Spearman menilai korelasi perbedaan viabilitas kelompok pasca-radiasi, dan pasca radiasi dan kultur. Uji Kruskall-Wallis menilai WPP kelompok pasca-radiasi antara masing-masing besar dosis. Uji Wilcoxon menilai WPP antara kelompok pre-radiasi dengan kelompok pasca-radiasi.Hasil: Waktu konfluensi kultur sel pasca radiasi rata-rata 4.33 hari. Rerata perbedaan viabilitas antara besar dosis radiasi tidak terdapat perbedaan yang bermakna secara statistik (p>0.05). Didapatkan korelasi positif viabilitas pasca radiasi dengan besar dosis radiasi namun tidak bermakna secara statistik (p>0.05) namun didapatkan korelasi negatif viabilitas pasca radiasi dan kultur dengan besar dosis radiasi dan bermakna secara statistik. Tidak didapatkan perbedaan bermakna median WPP antara kelompok pre-radiasi dan pasca-radiasi (p>0.05) dan perbedaan WPP diantara kelompok pasca radiasi (p>0.05).Kesimpulan: Tidak terdapat perbedaan secara statistic viabilitas dan WPP SPM jaringan adiposa pasca pajanan radiasi sinar-x c-arm sampai sampai dosis radiasi 32.34 mSv.

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ABSTRACTIntroduction. The use of adipose tissue derived mesenchymal stem cells (MSCs)in tissue engineering require implantation procedure that is safe and free of contamination. Minimally invasive procedure in the case of spinal cord injury using a c-arm device for MSC implantation causes x-ray exposure to the implanted cells. Viability and population doubling time (PDT) is a major component of the success of the implantation procedure. This study aims to determine the effect of c-arm x-ray exposure on MSC viability and PDT.Methods. This was an experimental study that used cryopreserved adipose tissue derived MSCs. Cells were thawed, propagated, and exposed to varying doses of c-arm x-ray radiation. Stem cell viability was measured, and then the cells were cultured to assess their PDT. Generalized linear models test was used to compare cell viability between post-thaw, post-propagation, post-radiation, post-culture post-radiation, and control and between radiation dose groups. Kruskal-Wallis test assessed PDT between various radiation doses in post-radiation groups. Wilcoxon test assessed PDT between pre-radiation and post-radiation groups.Results. Mean confluence period of adipose MSCs post radiation was 4.33 days. There was no statistically significant difference in MSC mean viability after exposure to x-ray radiation between each group and control (p>0.05). There was no significant positive correlation between post radiation viability and radiation dose (p > 0.05), however, there was significant negative correlation between post radiation post culture viability and radiation dose. There were no significant differences in PDT between pre- and post-culture post radiation groups and between various radiation doses in post-radiation groups (p>0.05).Conclusion. No statistical differences in MSC viability and PDT after x-ray radiation exposure of c-arm up to 32.34 mSv;Introduction. The use of adipose tissue derived mesenchymal stem cells (MSCs)in tissue engineering require implantation procedure that is safe and free of contamination. Minimally invasive procedure in the case of spinal cord injury using a c-arm device for MSC implantation causes x-ray exposure to the implanted cells. Viability and population doubling time (PDT) is a major component of the success of the implantation procedure. This study aims to determine the effect of c-arm x-ray exposure on MSC viability and PDT.Methods. This was an experimental study that used cryopreserved adipose tissue derived MSCs. Cells were thawed, propagated, and exposed to varying doses of c-arm x-ray radiation. Stem cell viability was measured, and then the cells were cultured to assess their PDT. Generalized linear models test was used to compare cell viability between post-thaw, post-propagation, post-radiation, post-culture post-radiation, and control and between radiation dose groups. Kruskal-Wallis test assessed PDT between various radiation doses in post-radiation groups. Wilcoxon test assessed PDT between pre-radiation and post-radiation groups.Results. Mean confluence period of adipose MSCs post radiation was 4.33 days. There was no statistically significant difference in MSC mean viability after exposure to x-ray radiation between each group and control (p>0.05). There was no significant positive correlation between post radiation viability and radiation dose (p > 0.05), however, there was significant negative correlation between post radiation post culture viability and radiation dose. There were no significant differences in PDT between pre- and post-culture post radiation groups and between various radiation doses in post-radiation groups (p>0.05).Conclusion. No statistical differences in MSC viability and PDT after x-ray radiation exposure of c-arm up to 32.34 mSv, Introduction. The use of adipose tissue derived mesenchymal stem cells (MSCs)in tissue engineering require implantation procedure that is safe and free of contamination. Minimally invasive procedure in the case of spinal cord injury using a c-arm device for MSC implantation causes x-ray exposure to the implanted cells. Viability and population doubling time (PDT) is a major component of the success of the implantation procedure. This study aims to determine the effect of c-arm x-ray exposure on MSC viability and PDT.Methods. This was an experimental study that used cryopreserved adipose tissue derived MSCs. Cells were thawed, propagated, and exposed to varying doses of c-arm x-ray radiation. Stem cell viability was measured, and then the cells were cultured to assess their PDT. Generalized linear models test was used to compare cell viability between post-thaw, post-propagation, post-radiation, post-culture post-radiation, and control and between radiation dose groups. Kruskal-Wallis test assessed PDT between various radiation doses in post-radiation groups. Wilcoxon test assessed PDT between pre-radiation and post-radiation groups.Results. Mean confluence period of adipose MSCs post radiation was 4.33 days. There was no statistically significant difference in MSC mean viability after exposure to x-ray radiation between each group and control (p>0.05). There was no significant positive correlation between post radiation viability and radiation dose (p > 0.05), however, there was significant negative correlation between post radiation post culture viability and radiation dose. There were no significant differences in PDT between pre- and post-culture post radiation groups and between various radiation doses in post-radiation groups (p>0.05).Conclusion. No statistical differences in MSC viability and PDT after x-ray radiation exposure of c-arm up to 32.34 mSv]"

"Pendahuluan. Sel punca mesenkimal merupakan jawaban untuk berbagai penyakit, termasuk orthopedi. Meskipun jumlah terbatas, prosedur invasif, nyeri, dan sel yang relatif sedikit, sumsum tulang masih menjadi sumber utama. Adiposa menjadi alternatif menjanjikan dengan kemampuan sebanding. Dengan meningkatnya harapan hidup, jumlah pasien tua meningkat dan menjadi sangat potensial untuk aplikasi sel punca. Namun, timbul kontroversi mengenai kualitas sel punca pada penuaan. Metode Penelitian. Penelitian dilakukan di Unit Pelayanan Terpadu Teknologi Kedokteran Sel Punca Rumah Sakit Cipto Mangunkusumo-Fakultas Kedokteran Universitas Indonesia, Jakarta sejak Oktober 2015 - Maret 2016. 12 subjek dibagi menjadi tiga kelompok usia; 15-30 tahun, 31-40 tahun, dan 41-55 tahun dan dilakukan pengambilan sumsum tulang krista iliaka posterior dan adiposa, kemudian dilakukan isolasi dan kultur sel punca mesenkimal. Peneliti melakukan analisis karakteristik biologis, waktu penggandaan populasi, diferensiasi osteogenik, dan pewarnaan Alizarin. Seluruh data dianalisis dengan SPSS 20. Temuan Penelitian. Karakteristik biologis dan pewarnaan Alizarin Red menunjukkan tidak ada perbedaan bermakna sel punca mesenkimal sumsum tulang dan adiposa pada kelompok usia sama(p>0,05). Waktu penggandaan populasi menunjukkan adanya perbedaan signifikan sel punca mesenkimal sumsum tulang dan adiposa pada kelompok 31-40 tahun(p=0,028) dan 41-55 tahun(p=0,035). Kesimpulan. Sel punca mesenkimal adiposa menunjukkan karakteristik biologis, waktu penggandaan populasi, dan diferensiasi osteogenik yang konstan. Sel punca mesenkimal sumsum tulang menunjukkan waktu penggandaan populasi yang menurun seiring usia, berbeda dengan karakteristik biologis dan diferensiasi osteogenik. Adiposa dapat menjadi pilihan sumber sel punca mesenkimal pada setiap golongan usia.

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Introduction. Mesenchymal stem cell is the answer of many medicine problems, including orthopaedic. Bone marrow is still the main source. Because of limited source, invasive procedure, pain, and relative less cell, adipose will be promising source with equal regenerating and differentiating ability. Along with increasing life expectancy, geriatric population is increasing as well as the potential need for stem cell application. Yet there is still controversy about stem cell quality in aging.

Methods. This study was conducted in Stem Cell Medical Technology Integrated Service Unit Cipto Mangunkusumo General Hospital-Faculty of Medicine Universitas Indonesia, Jakarta, October 2015 - March 2016. 12 patients were divided into 3 age group; 15-30 year, 31-40 year, and 41-55 year. Bone marrow from posterior iliac crest and adipose tissue were collected, mesenchymal stem cell isolation and culture were done subsequently. Biological characterization, Population Doubling Time, osteogenic differentiation, and alizarin red assay were carried out. All data was analyzed using SPSS 20.

Results. No significant difference was observed in biological characteristic and Alizrin red assay of bone marrow and adipose mesenchymal stem cell among age group (p>0.05). There is significant difference in Population Doubling time in 31- 40 year group(p=0.000) and 41-55 year group(p=0.000).

Conclusions. Adipose mesenchymal stem cell had steady biological characteristic, Population Doubling Time, and osteosteogenic differentiation. Bone marrow mesenchymal stem cell had increasing population doubling time in increasing age, apart from biological characteristic and osteogenic differentiation. Adipose could be the source of choice in harvesting mesenchymal stem cell at any age."

"Pendahuluan: Penelitian in vitro menggambarkan inferioritas osteogenesis SPM adiposa dibandingkan dengan SPM sumsum tulang. Sebaliknya, penelitian in vivo menunjukkan kemiripan potensi osteogenik keduanya. penelitian ini mencoba mengetahui perbedaan kapasitas osteogenik antara keduanya dengan mengukur ekspresi Bone Morphogenetic Protein (BMP)-2 dan BMP Reseptor II, juga proses penyembuhan tulang dengan pengukuran histomorfometri.Metode: Delapan belas tikus Sprague dawley (SD) dilakukan defek tulang femur 5mm. Tikus dibagi tiga kelompok yang terdiri dari kontrol, implantasi SPM sumsum tulang + Hydroxypatite, dan implantasi SPM adiposa + Hydroxypatite. Tikus dikorbankan pada minggu kedua kemudian penilaian histomorfometri kuantitatif dilakukan dengan Image-J. Paramater yang diukur adalah luas total kalus, % area penulangan, % area kartilago, dan % area fibrosis. Dilakukan penilaian imunohistokimia menggunakan intensitas pewarnaan dan skor Imunoreaktivitas (IRS).Hasil: Kelompok SPM sumsum tulang menunjukkan ekspresi BMPR II lebih tinggi dibandingkan kelompok lainnya. Ekspresi BMPR II dianalisis dan didapatkan hasil yang signifikan (p= 0,04) dengan median 4.00 ± 2.75. Kelompok SPM sumsum tulang dan adiposa juga menunjukkan proses penyembuhan tulang yang lebih baik dibandingkan kelompok kontrol (p = 0,001). Tidak ada perbedaan yang signifikan antara SPM sumsum tulang dan SPM adiposa yang diukur pada % total area kalus (p = 1.000),% area penulangan (p = 1.000),% kartilago (p = 0,493) dan % fibrosis (p = 0,128).Diskusi: SPM adiposa memiliki kemampuan penyembuhan tulang yang serupa dengan SPM sumsum tulang. Growth factor dan reseptornya penting namun bukan satu-satunya faktor penyembuhan tulang.

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Introduction: In vitro studies describe inferior osteogenesis of Adiposes to Bone Marrow Mesenchymal Stem Cell (MSC). Contrary, in vivo studies showing the resemblance of osteogenic potential between both groups. This study tries to investigate the difference of osteogenic capacity between BMSCs and ASCs by quantifying the expression of Bone Morphogenetic Protein (BMP)-2 and BMP receptor (BMPR) II also the bone healing process by histomorphometry measurement.Methods: Eighteen Sprague dawley (SD) rats were induced with 5mm femoral bone defect, then divided into three groups that consist of Control, Implementation of BMSC+Hydroxypatite, and Implementation of ASC+Hydroxypatite. They were sacrificed after 2 weeks, then performed histomorphometry assessment with Image-J. The measured paramater were total area of callus, % of osseous area, % of cartilage area, and % of fibrotic area. The immunohistochemistry measurement performed by staining intensity and immunoreactivity score (IRS).Results: The BMSC group showed higher expression of BMPR II compare to others. The expression of BMPR II was analyzed statistically and showed significant result (p=0.04) with median 4.00 ± 2.75. Both BMSC and ASC group have significantly better bone healing process compared with control group (p=0,001). There are no significant differences between ASC and BMSC measured in %total callus area (p=1.000), %Osseous area (p=1.000), %Cartilage area (p=0.493) and % Fibrous area (p=0.180). Discussions: ASC bone healing ability are similar to BMSC. Growth factor and its receptor are important but not sole contributing factor for bone healing."

"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

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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"

"Latar Belakang : Diferensiasi sel punca mesenkimal (SPM) menjadi osteoblas dan pertumbuhannya pada lingkungan mikroskopis yang terpajan debris bakteri Mycobacterium tuberculosis secara in vitro tidak menunjukkan gangguan berarti. SPM memiliki potensi imunomodulator dan membantu memperbaiki jaringan yang rusak. Penelitian ini bertujuan untuk mendapatkan pemahaman mengenai manfaat SPM pada eradikasi infeksi, pembentukan tulang dan fusi lesi tulang belakang.Metode : Penelitian ini merupakan penelitian eksperimental pada hewan kelinci yang dilaksanakan dalam 2 tahap. Pada tahap pertama dua puluh tujuh ekor kelinci diinokulasi bakteri Mycobacterium tuberculosis pada korpus vertebra T12. Pengamatan dilakukan terhadap berat badan, suhu badan, populasi Th1, Th2 dan rasio Th1/Th2, keberadaan bakteri serta reaksi jaringan. Pada tahap kedua kelinci yang diinokulasi bakteri Mycobacterium tuberculosis dijadikan sebagai sampel dan dilakukan prosedur tata laksana total Subroto Sapardan, penambahan skafold, penambahan SPM dan pemberian obat anti tuberkulosis. Dengan mengeluarkan kelinci yang tidak memenuhi syarat diperoleh masing-masing 7 kelinci kelompok transplantasi SPM dan kelompok kontrol. Pengamatan dilakukan terhadap berat badan, suhu badan, populasi Th1, Th2 dan rasio Th1/Th2, keberadaan bakteri, reaksi jaringan, ekspresi CBFA-1, sekresi OPN, sekresi ALP, hitung osteoblas, hitung osteosit, kadar kalsium lesi, pembentukan tulang per mm2 defek, dan uji pergerakan tulang.Hasil : Pada tahap pertama diperoleh 100 % kelinci spondilitis tuberkulosis berdasarkan pemeriksaan histopalogi. Pada tahap kedua diperoleh persentase normalisasi pemeriksaan BTA positif pada kelompok SPM (1/1) lebih banyak dibandingkan kelompok kontrol (1/2). Persentase pemeriksaan ALP positif pada kelompok SPM (7/7) lebih banyak dibandingkan kelompok kontrol (5/7). Rerata pembentukan tulang per mm2 defek pada kelompok SPM (1,98 mm2) lebih besar dibandingkan kelompok kontrol (0,88 mm2) (p<0,05). Persentase kelinci yang mengalami fusi pada kelompok SPM (29 %) lebih banyak dibandingkan kelompok kontrol (0 %).Simpulan : Transplantasi SPM ke dalam defek lesi spondilitis tuberkulosis meningkatkan eradikasi infeksi, terbentuknya tulang baru dan capaian fusi tulang belakang.

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Backgrounds: Mesenchymal stem cell (MSC) differentiation and growth to osteoblast in micro environment exposed with Mycobacterium tuberculosis debris did not show significant effect in vitro. MSC has immunomodulatory potency and helps repairing damaged tissues. This research aims to understand MSC benefits on infection eradication, bone formation and spinal lesion fusion.

Methods: Two steps of experimental research were done using rabbit as a model on this research. At the first step, twenty seven rabbits were inoculated with Mycobacterium tuberculosis on T12 vertebral body. Rabbit's weight, temperature, Th1 and Th2 population with Th1/Th2 ratio, bacteria's existence, and tissue reactions were examined. On the second step, the rabbits previously inoculated with Mycobacterium tuberculosis were used. Rabbits were not eligible for second step experimental were excluded and 7 rabbits were finally used for each MSC transplantation group and the control group. Observation on the weight, temperature, Th1 and Th2 population with Th1/Th2, bacteria's existence, tissue reactions, core binding factor alfa -1 (CBFA-1)expression, osteopontin (OPN) secretion, alkaline phosphatase (ALP) secretion, osteoblast count, osteocytes count, calcium intralesion level, bone formation per milimeter square defect, and bone movement test were done.

Results: On the first step, 100 % rabbits with spondylitis tuberculosis were yielded based on positive histologic test. On the second step, positive percentage on Acid Fast Bacilli (AFB) test was higher on MSC group (1/1) compared to control group (1/2). Positive ALP percentage on MSC group was also higher (7/7) than control group (5/7). Mean bone formation per milimeter square of defect on the MSC group (1.98 mm2) was larger than the control group (0.88 mm2) (p<0.05). Number of rabbit underwent fusion were higher in the MSC group (29 %) than the control group (0 %).

Conclusion: MSC transplantation on spondylitis tuberculosis lesion defect could increase the eradication of infection, new bone formation and spinal fusion outcome"

"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.

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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."

"ABSTRAKPendahuluan. Penggunaan autograft dalam fusi interkorpus tulang belakang masih menjadi pilihan utama, tetapi jumlah yang terbatas dan morbiditas pada tempat donor mendorong penggunaan substitusi tulang. Kombinasi HA dan DBM menjadi pilihan utama selain autograft dalam fusi interkorpus, namun hasil yang ada menunjukan variasi diantara jenis DBM. Tujuan penelitian ini untuk mengevaluasi luaran klinis dan radiologis fusi interkorpus lumbal menggunakan kombinasi DBM dan HA.Metode Penelitian. Penelitian ini merupakan prospektif cohort pada 35 pasien yang terbagi atas 18 pasien kelompok autograft dan 17 pasien kelompok kombinasi HA dan DBM. Pasien merupakan pasien spondilosis lumbal yang diindikasikan untuk tindakan operatif. Evaluasi klinis pada masing-masing kelompok pasca operasi menggunakan VAS, JOA dan ODI yang dinilai pada bulan ke-3, 6 dan 12. Evaluasi radiologis pada masing-masing berupa fusi di evaluasi dengan ct scan pada bulan ke-12. Karakteristik pasien seperti jenis kelamin, usia, riwayat merokok, level operasi, dan BMI juga dievaluasi.Temuan Penelitian. Dua orang ahli bedah orthopaedi tulang belakang melakukan operasi stabilisasi posterior dan TLIF. Terdapat 55 pasien (27 kelompok autograft, 28 kelompok kombinasi HA dan DBM) yang masuk ke dalam kriteria, 9 pasien dari masing-masing kelompok di eksklusi karena tidak dapat di follow up sampai 12 bulan. Perbandinagn skor VAS, JOA dan ODI diantara kedua kelompok tidak menunjukan perbedaan yang bermakna dengan nilai p masing-masing 0,599, 0,543 dan 0,780. Perbandingan fusi antara kelompok autograft dan kombinasi HA dan DBM menunjukan nilai p 1,000, sehingga tidak bermakna secara statistik.Simpulan Hasil luaran klinis dan radiologis pada penggunaan kombinasi HA dan DBM dalam fusi interkorpus tidak menunjukan inferioritas bila dibandingkan dengan autograft. Kombinasi HA dan DBM dapat dipertimbangkan sebagai alternatif bagi pasien spondilosis lumbal yang diindikasikan untuk tindakan operatif.

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ABSTRACTIntroduction The use of autograft still remains a gold standard in lumbar interbody fusion surgery, but the limited amount and donor site morbidity encourages the use of bone substitute. Combination of HA and DBM become a main choice other than autograft in lumbar interbody fusion, however there were variable result between DBM product. These research was aimed to evaluate the clinical and radiological outcome of interbody fusion using combination of DBM and HA.Methods A cohort prospective research was conducted in 35 patients that were divided into 18 autograft group patients and 17 combination of HA and DBM group patient. All the patients were diagnosed with lumbar spondylosis and indicated for surgery. Clinical evaluation on each group was evaluated using VAS, JOA and ODI on the 3rd, 6th and 12th month post operatively. Radiologic outcome of fusion was evaluated using ct scan on the 12th month. Other patient characteristic such as sex, age, smoking history, level operation dan BMI were also evaluated in this research.Results Two orthopaedic spine surgeon conducted the posterior stabilization and TLIF procedure. There were 55 patients (27 autograft group patients, 28 combination of HA and DBM group patients) that was included according to the criteria, 9 patients on each group were excluded due to loss of follow up below 12 months. Comparison of VAS, JOA and ODI score between the two group did not show any difference that significant statiscally with the p value was 0.599, 0.543, and 0.780 each. Comparison of fusion rate between the two groups showed p value of 1.000 which was not significant statiscally.Conclusions Clinical and radiological outcome of combination of HA and DBM in lumbar interbody fusion did not show inferiority compared with autograft. Combination of HA and DBM can be considered as an alternative in lumbar spondylosis patient that need operative procedure"

"ABSTRAKPendahuluan. Penggunaan autograft dalam fusi interkorpus tulang belakangmasih menjadi pilihan utama, tetapi jumlah yang terbatas dan morbiditas padatempat donor mendorong penggunaan substitusi tulang. Kombinasi HA dan DBMmenjadi pilihan utama selain autograft dalam fusi interkorpus, namun hasil yang ada menunjukan variasi diantara jenis DBM. Tujuan penelitian ini untukmengevaluasi luaran klinis dan radiologis fusi interkorpus lumbal menggunakankombinasi DBM dan HA.Metode Penelitian. Penelitian ini merupakan prospektif cohort pada 35 pasienyang terbagi atas 18 pasien kelompok autograft dan 17 pasien kelompokkombinasi HA dan DBM. Pasien merupakan pasien spondilosis lumbal yangdiindikasikan untuk tindakan operatif. Evaluasi klinis pada masing-masingkelompok pasca operasi menggunakan VAS, JOA dan ODI yang dinilai padabulan ke-3, 6 dan 12. Evaluasi radiologis pada masing-masing berupa fusi dievaluasi dengan ct scan pada bulan ke-12. Karakteristik pasien seperti jeniskelamin, usia, riwayat merokok, level operasi, dan BMI juga dievaluasi.Temuan Penelitian. Dua orang ahli bedah orthopaedi tulang belakang melakukan operasi stabilisasi posterior dan TLIF. Terdapat 55 pasien (27 kelompok autograft, 28 kelompok kombinasi HA dan DBM) yang masuk ke dalam kriteria, 9 pasien dari masing-masing kelompok di eksklusi karena tidak dapat di follow up sampai 12 bulan. Perbandinagn skor VAS, JOA dan ODI diantara kedua kelompok tidak menunjukan perbedaan yang bermakna dengan nilai p masing-masing 0,599, 0,543 dan 0,780. Perbandingan fusi antara kelompok autograft dan kombinasi HA dan DBM menunjukan nilai p 1,000, sehingga tidak bermakna secara statistik.Simpulan Hasil luaran klinis dan radiologis pada penggunaan kombinasi HA danDBM dalam fusi interkorpus tidak menunjukan inferioritas bila dibandingkandengan autograft. Kombinasi HA dan DBM dapat dipertimbangkan sebagaialternatif bagi pasien spondilosis lumbal yang diindikasikan untuk tindakanoperatif.

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ABSTRACTIntroduction The use of autograft still remains a gold standard in lumbarinterbody fusion surgery, but the limited amount and donor site morbidityencourages the use of bone substitute. Combination of HA and DBM become amain choice other than autograft in lumbar interbody fusion, however there were variable result between DBM product. These research was aimed to evaluate the clinical and radiological outcome of interbody fusion using combination of DBM and HA.Methods A cohort prospective research was conducted in 35 patients that were divided into 18 autograft group patients and 17 combination of HA and DBM group patient. All the patients were diagnosed with lumbar spondylosis and indicated for surgery. Clinical evaluation on each group was evaluated using VAS, JOA and ODI on the 3rd, 6th and 12th month post operatively. Radiologic outcome of fusion was evaluated using ct scan on the 12th month. Other patient characteristic such as sex, age, smoking history, level operation dan BMI were also evaluated in this research.Results Two orthopaedic spine surgeon conducted the posterior stabilization and TLIF procedure. There were 55 patients (27 autograft group patients, 28combination of HA and DBM group patients) that was included according to the criteria, 9 patients on each group were excluded due to loss of follow up below 12 months. Comparison of VAS, JOA and ODI score between the two group did not show any difference that significant statiscally with the p value was 0.599, 0.543, and 0.780 each. Comparison of fusion rate between the two groups showed p value of 1.000 which was not significant statiscally.Conclusions Clinical and radiological outcome of combination of HA and DBMin lumbar interbody fusion did not show inferiority compared with autograft.Combination of HA and DBM can be considered as an alternative in lumbarspondylosis patient that need operative procedure"

"Defek kritis tulang panjang adalah kondisi bagian tulang yang hilang dengan ukuran lebih dari 2 cm dan atau 50% diameter tulang, sehingga sulit untuk mengalami regenerasi. Salah satu penatalaksanaan yang dapat dilakukan adalah tindakan transplantasi autologous namun peningkatan risiko morbiditas pada pendonor menyebabkan diperlukannya tata laksana alternatif untuk defek kritis tulang panjang. Penggunaan eksosom sel punca mesenkimal adiposa atau PRF telah menunjukkan hasil yang menjanjikan pada penelitian sebelumnya. Kami bertujuan untuk mengevaluasi efek penggunaan eksosom sel punca mesenkimal adiposa dan PRF terhadap defek kritis tulang panjang pada tikus Sprague-Dawley. Penelitian ini merupakan penelitian quasi eksperimental post-test only control group design pada hewan coba tikus Sprague Dawley. Sampel diambil secara acak dari tikus putih spesies Sprague Dawley jantan yang berusia 8-12 minggu, dengan berat sekitar 250 – 350 gram. Sebanyak 30 ekor tikus dibagi ke dalam 5 kelompok, yaitu kelompok perlakuan hidroksiapatit (HA) dan bone graft (BG) (kelompok I), kelompok perlakuan HA, BG, dan PRF (kelompok II), kelompok perlakuan HA, BG dan eksosom sel punca mesenkimal adiposa (kelompok III), kelompok perlakuan HA, BG, PRF, dan eksosom sel punca mesenkimal adiposa (kelompok IV), dan kelompok perlakuan HA, PRF, dan eksosom sel punca mesenkimal adiposa (kelompok V). Setiap tikus kemudian dibuat defek tulang femur sebesar 5mm yang difiksasi interna menggunakan K-wire ukuran 1,4 mm. Histomorfometri dan BMP-2 dilakukan untuk menilai proses penyembuhan tulang pada setiap kelompok perlakuan. Pada analisis RT-PCR, kelompok IV (HA + BG + eksosom sel punca mesenkimal adiposa) memiliki ekspresi gen BMP-2 tertinggi dibandingkan dengan kelompok lainnya. Sebaliknya, kelompok III (HA + BG + eksosom sel punca mesenkimal adiposa + PRF) memiliki tingkat chordin tertinggi dibandingkan dengan kelompok lainnya. Secara umum, kelompok yang diintervensi dengan eksosom sel punca mesenkimal adiposa atau PRF memiliki ekspresi BMP-2 yang lebih tinggi dibandingkan dengan kontrol. Namun, kami tidak menemukan perbedaan yang signifikan antar kelompok dalam analisis statistik. Pembentukan jaringan ikat pada penyembuhan tulang predominan dibandingkan pembentukan jaringan tulang untuk semua kelompok. Kelompok dengan pemberian kombinasi eksosom sel punca mesenkimal adiposa, PRF, HA menunjukkan hasil yang setara/ekuivalen dengan HA+ BG. Dalam penelitian ini, penggunaan eksosom sel punca mesenkimal adiposa dan/atau PRF telah menunjukkan peningkatan aktivitas osteogenic yang ditunjukkan dengan peningkatan laju penyembuhan tulang. Kuantifikasi BMP-2 dapat menunjukkan aktivitas osteogenic pada tikus yang ditatalaksana dengan eksosom sel punca mesenkimal adiposa, bone graft dan HA. Selain itu, penggunaan eksosom sel punca mesenkimal adiposa yang dikombinasikan dengan PRF menunjukkan efek yang saling mendukung. Hal ini tampak pada kombinasi eksosom sel punca mesenkimal adiposa, PRF, HA tanpa BG menunjukkan hasil yang setara/ekuivalen dengan HA+ BG. Hasil histomorfometri menunjukkan aktivitas osteogenic yang baik pada tikus yang ditatalaksana dengan eksosom sel punca mesenkimal adiposa dan/atau PRF. Namun, efek ini tidak terlalu tampak pada kombinasi eksosom sel punca mesenkimal adiposa, PRF, HA dan BG meskipun hasil ini memiliki tren yang lebih tinggi dibandingkan dengan kelompok kontrol. Hal ini mendukung sinergi antara eksosom sel punca mesenkimal adiposa dan PRF. Penggunaan PRF dan eksosom sel punca mesenkimal adiposa memiliki luaran histomorfometri dan molekular (BMP-2 dan Chordin) yang sebanding dengan penggunaan bone graft pada defek tulang kritis pada tikus Sprague Dawley.

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Critical long bone defects is defined as a defect of over 2 cm or 50% of the bone diameter that leads to a small chance of healing. Autologous bone graft has been proposed as a treatment for critical bone defects in prior studies. However, unreliable results and donor-site morbidity call for an alternative treatment in critical long bone defect. Biological augmentation with ASCs exosome and PRF has shown promising results in bone regeneration in prior studies. We aimed to evaluate the efficacy of ASCs exosome and PRF in treating critical long bone defect in Sprague-Dawley rats. This study was a quasi-experimental post-test only control group design on Sprague-Dawley rats. Samples were taken randomly from male Sprague-Dawley white rats aged 8 to 12 weeks, weighing approximately 250 to 350 grams. A total of 30 rats were divided into 5 groups: hydroxyapatite (HA) and bone graft (BG) treatment group (group I); HA, BG, and PRF treatment group (group II); HA, BG, PRF and ASCs exosome treatment group (group III); HA, BG, and ASCs exosome treatment group (group IV); and HA, PRF, and ASCs exosome treatment group (group V). A 5 mm femur bone defect was created that was internally fixed using a 1.4 mm K-wire threaded. Histomorphometry and BMP-2 was performed to evaluate bone healing process in each group. On RT-PCR analysis, group IV (HA+BG+ASCs exosome) had the highest BMP-2 gene expression compared to other groups. In the contrary, group III (HA+BG+ASCs exosome+PRF) has the highest chordin level compared to other groups. In general, the group intervened by ASCs exosome or PRF has a higher BMP-2 expression compared to control. However, we did not find any significant difference between groups in statistical analysis. Histomorphometry examination showed increased bone healing progression in groups with ASCs or PRF. The use of biological augmentation to increase the speed and rate of bone healing especially in critical bone defect has been shown in previous study. In this study, the use of ASCs exosome and/or PRF has shown increased osteogenic activities that translates into increased rate of bone healing. The quantification of BMP-2 could show the osteogenic activities in rats treated with ACSs exosome with BG and HA. In addition, the use of adipose mesenchymal stem cell exosomes in combination with PRF showed a mutually supportive effect. This was seen in the combination of adipose mesenchymal stem cell exosomes, PRF, HA without BG showed equivalent results with HA + BG. Histomorphometric results showed good osteogenic activity in rats treated with adipose mesenchymal stem cell exosomes and/or PRF. However, this effect was less pronounced in the combination of adipose mesenchymal stem cell exosomes, PRF, HA and BG although this result had a higher trend compared to the control group. This supports the synergy between adipose mesenchymal stem cell exosomes and PRF. The ASCs exosome showed a positive effect on osteogenesis in critical long bone defects in Sprague-Dawley rats."