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Abstrak :
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. ......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

Abstrak :
ABSTRAK Latar Belakang: Sel punca CD34+ adalah sel punca hematopoietik yang positif terhadap penanda sel CD34 dan mempunyai potensi regenerasi. Potensinya dapat ditingkatkan dengan penambahan asam hialuronat dan faktor pertumbuhan. Tujuan penelitian adalah menghasilkan regenerasi tulang rawan hialin pada defek osteokondral sendi lutut tikus Spraque Dawley (SD) non rekayasa melalui penyuntikan intraartrikular sel punca CD34+ darah tepi manusia, asam hialuronat, TGF-β1, IGF, FGF dan Fibronektin. Metode: Penelitian dilakukan 3 tahap. Tahap 1: Pembuatan Model Defek, Model Intervensi, dan Uji Toksik. Tahap 2: Penyiapan Sel dan Pembuatan Suspensi. Tahap 3: Intervensi. Pada tahap intervensi, 30 tikus SD dibagi menjadi 3 kelompok dan setiap tikus dibuat defek dangkal dan dalam pada sendi lutut. Setelah luka operasi ditutup, tiap tikus diberi suspensi secara intraartrikular. Kelompok kontrol diberi PBS, kelompok perlakuan 1 diberi sel CD34+, kelompok perlakuan 2 diberi sel CD34+, asam hialuronat, TGF-β1, IGF, FGF dan Fibronektin. Setiap kelompok dievaluasi laboratoris, radiologis, makroskopis dan mikroskopis pada minggu ke-4 dan ke-8. Hasil penelitian diuji secara statistik (Uji Manova). Hasil : Tidak terjadi reaksi penolakan. Terjadi perbedaan bermakna antara kelompok perlakuan dengan kelompok kontrol dalam kadar Hb (p = 0,016), Trombosit (p = 0,009), SGPT (p = 0,000), dan Kreatinin (p = 0,029), namun mikroskopis hati dan ginjal normal. Pemberian sel CD34+ tidak memperbaiki skor radiologis (p = 0,074), namun terjadi regenerasi, skor makroskopis defek dangkal (p = 0,000), makroskopis defek dalam (p = 0,000), mikroskopis defek dangkal (p = 0,000) dan mikroskopis defek dalam (p = 0,000). Kelompok perlakuan 2 tidak berbeda dengan kelompok perlakuan 1, skor makroskopis defek dangkal (p=1,000), mikroskopis defek dangkal (p = 1,000) dan defek dalam (p = 0,818), namun perlakuan 2 lebih baik dari perlakuan 1 pada makroskopis defek dalam (p = 0,023). Skor defek dangkal dan defek dalam tidak berbeda bermakna pada kelompok kontrol dan perlakuan pada minggu ke-4 dan ke-8 (p makroskopis = 0,793, p mikroskopis = 0,754). Skor minggu ke-4 dan ke-8 tidak berbeda bermakna pada kelompok kontrol dan perlakuan, pada defek dangkal dan defek dalam (p radiologis = 0,200, p makroskopis dangkal = 0,507, makroskopis dalam = 0.350, p mikroskopis dangkal = 0,446, p mikroskopis dalam = 0,239). Simpulan : Sel punca CD34+ darah tepi manusia dapat menghasilkan regenerasi hialin pada model defek osteokondral. Penambahan asam hialuronat dan faktor pertumbuhan tidak meningkatkan hasil regenerasi. Pembuatan mikrofraktur pada defek osteokondral tidak meningkatkan hasil regenerasi. Hasil regenerasi minggu ke- 8 tidak lebih baik dari minggu ke-4.

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ABSTRACT Background: CD34+ is hematopoietic stem cell that is positive to CD34 cell markers and potential for tissue regeneration. The regeneration potential for the cartilage has never been researched. The potential can be increased by adding the hyaluronic acid and growth factors. The research was aimed at producing the hyaline cartilage regeneration in the osteochondral defect of naïve Spraque Dawley (SD) rats? knee joints by intraartricularily injecting human?s peripheral blood CD34+ stem cell, hyaluronic acid and TGF-β1, IGF, FGF, and Fibronectin. Methods: The research comprised 3 stages. Stage 1: The Development of Defect Models, Intervention Model, and Toxic Test. Stage 2: Preparation of Cells and Suspension Making. Stage 3: Intervention. During the intervention process, 30 SD rats were grouped into three groupsandthesuperficialanddeepdefectsweremadeontheirknees. Afterthesurgicalwound was covered, each rat was intraartricularily injected by suspension. The control group received PBS, the treatment group 1 received CD34+ cell, the treatment group 2 was given CD34+ cell, hyaluronic acid, and TGF-β1, IGF, FGF, and Fibronectin. Every group was evaluated in the laboratory, radiologically, macroscopically, and microscopically on the 4th and 8th weeks. The research result was analyzed statistically (Manova Test). Result: There was no rejection. There were significant differences between the treatment group and control group with respect to the Hb (p = 0.016), thrombocyte (p = 0.009), SGPT (p = 0.000), and creatinine (p = 0.029), but the liver and kidney microscopic were normal. The administration of CD34+ cells did not improve the radiological score (p = 0.074), but there was regeneration, the macroscopic score of the superficial defect (p=0.000), macroscopic score of the deep defect (p=0.000), microscopic score of the superficial defect (p=0.000), and microscopic score of the deep defect (p=0.000). The macroscopic score of the superficial defect of the treatment group 2 was not significantly different from the results of the treatment group 1 (p=1.000), macroscopic score of the superficial defect (p=1.000), microscopic score of the superficial defect (p=1.818), and the microscopic score of the deep defect (p=0.023). There were no significant differences between the score of the superficial defect and score of the deep defect in the control and treatment groups in the 4th and 8th weeks (p macroscopic = 0.793, p microscopic = 0.754). There were no differences with respect to the scores in the 4th and 8th weeks in the control and treatment groups and with respect to the superficial and deep defects (p radiological = 0.200, p superficial macroscopic = 0.507, deep macroscopic = 0.350, p superficial microscopic = 0.446, p deep microscopic = 0.239). Conclusion: Human?s peripheral blood CD34+ stem cell can produce hyaline regeneration in the osteochondral defect models. The addition of hyaluronic acid and growth factors does not improve the regeneration results. The microfracture deep in the osteochondral defect does not improve the regeneration result. The regeneration result in the 8th week is better than the result in the 4th week.