Please use this identifier to cite or link to this item:
https://ptsldigital.ukm.my/jspui/handle/123456789/515831Full metadata record
| DC Field | Value | Language |
|---|---|---|
| dc.contributor.advisor | Ruszymah Idrus, Prof. Dr. | |
| dc.contributor.author | Ude Chinedu Cletus (P51900) | |
| dc.date.accessioned | 2023-10-16T09:31:25Z | - |
| dc.date.available | 2023-10-16T09:31:25Z | - |
| dc.date.issued | 2014-05-08 | |
| dc.identifier.other | ukmvital:80557 | |
| dc.identifier.uri | https://ptsldigital.ukm.my/jspui/handle/123456789/515831 | - |
| dc.description | Osteoarthritis (OA) is a major degenerative joint disease and to date there is no definitive cure for it. Cell based therapies hold some promises as treatment options. Adipose stem cells (ADSC) and bone marrow stem cells (BMSC) are multipotent cells with the potentials for cartilage regeneration. The aims of this study are: to evaluate the lesions developed in a surgically induce osteoarthritis on sheep knee via arthroscopy; to evaluate the effectiveness of the intra-articular injection of autologous chondrogenic induced ADSC and BMSC; to evaluate the homing of the injected chondrogenic induced cells within the knee joint using PKH26 dye; to compare the effectiveness of using TGFβ-3 alone and in combination with BMP-6 in hyaline cartilage formation; and lastly to assess the functions of the OA knee joints via electromyography and the mechanical qualities of the regenerated cartilages via compression modulus after chondrogenic induced cell treatments. Osteoarthritis was induced on the right knee joint of 30 sheep: 9 as control, 12 ADSC and 9 BMSC samples by complete resection of the anterior cruciate ligament and medial meniscus, followed by a 3-weeks exercise regimen. ADSC and BMSC were harvested from the infra-patella fat pad and the iliac crest respectively. Stem cells from experimental sheep were culture expanded and induced to chondrogenic lineage using TGF-β3 as main induction factor and subsequently optimized with BMP-6. Test sheep received single dose of 2x107 autologous PKH26-labeled, chondrogenic induced ADSC or BMSC as 5mls injections, while controls received 5mls basal medium. Sheep were sacrificed 6 weeks post cell injection. Grossly, treated knee joints showed regenerated de novo cartilages. On the ICRS grade scores, ADSCs and BMSCs groups had significantly lower scores than controls (P<0.05). Histological staining revealed loosely packed matrixes of de novo cartilages and immunostaining demonstrated the presence of cartilage specific proteins. PKH26 dye fluorescence on the resected portions of regenerated neo-cartilages revealed the homing of the injected cells. With the addition of BMP-6, BMSCs had higher expression of Collagen I, and SOX9, while ADSCs had higher expression of Collagen II and Aggrecan genes (P<0.05). TGF-β3 alone favoured BMSCs chondrogenesis; while the combination of BMP-6 increased chondrogenic gene expressions on both, with greater enhancement on ADSCs. Posttreatment EMG showed that the hind limbs shifted their weight back towards the right; returning to Pre-OA values in both cell treatment groups (p<0.05). The mechanical properties of our engineered cartilages on the different regions were comparable with the native cartilage in compression modulus and strength. Our tissue engineered cartilages possessed evidences of functional correlation to their structural regenerations. Autologous ADSCs and BMSCs could be promising candidates for cartilage regeneration in osteoarthritis.,Osteoarthritis (OA) adalah sejenis penyakit sendi degeneratif dan terapi berasaskan sel memberi harapan dalam mengubati penyakit ini. Sel stem adipos (ADSC) dan sumsum tulang (BMSC) adalah sel multipoten yang berpotensi untuk regenerasi tisu rawan. Tujuan kajian ini adalah untuk menganalisa kerosakan rawan dengan OA yang diaruh pada model kambing biri-biri menggunakan teknik arthroskopi; mengkaji keberkesanan suntikan intra-artikular sel ADSC dan BMSC yang diaruh kepada leluhur kondrosit; menganalisa kehadiran sel yang disuntik di sendi lutut, menggunakan pewarnaan PKH26 ; membandingkan keberkesanan penggunaan TGFβ-3 sahaja dan dengan kombinasi BMP-6 dalam pembentukan rawan hialin dan akhir sekali, untuk menilai fungsi sendi lutut melalui elektromyography dan ciri mekanikal rawan yang terjana selepas rawatan dengan sel teraruh. Pengaruhan kepada OA telah dilakukan pada sendi lutut kanan 30 ekor kambing biri-biri: 9 kawalan, 12 ADSC dan 9 BMSC dengan membuang ligamen krusiate anterior dan meniskus medial, diikuti dengan 3 minggu regimen senaman. ADSC dan BMSC masing-masing diperolehi dari pad infra-patela dan krista iliak kambing biri-biri. Sel stem tersebut telah dikultur dan diaruh kepada leluhur kondrogenik menggunakan TGF-β3 sebagai faktor induksi utama dan kemudiannya bersama BMP-6. Kambing biri-biri telah menerima suntikan 5ml yang terdiri daripada 2x107 sel kondrosit teraruh daripada ADSC dan BMSC berlabel dengan PKH26, manakala kumpulan kawalan menerima 5ml medium basal. Kambing biri-biri dikorbankan 6 minggu selepas suntikan sel. Pemerhatian secara mata kasar menunjukkan regenerasi rawan pada sendi lutut yang diuji. Berdasarkan kepada pemarkahan ICRS, kumpulan ADSC dan BMSC mempunyai skor yang jauh lebih rendah berbanding kawalan (p<0.05). Pewarnaan histologi pula menunjukkan wujudnya matriks pra-matang pada tisu rawan yang terbentuk manakala pewarnaan imuno menunjukkan protein khusus rawan. Pewarnaan positif PKH26 pada tisu rawan baru menunjukkan ianya terbentuk daripada sel yang disuntik. Penambahan BMP-6meningkatkan kolagen 1 dan SOX-9 pada BMSC manakala peningkatan kolagen 2 dan aggrecan pada ADSC (p<0.05). TGF-β3 sahaja memihak kepada kondrogenesis BMSC, manakala gabungan dengan BMP-6 meningkatkan pengekspresan gen kondrogenik pada kedua-duanya, dengan peningkatan yang lebih pada ADSC. Analisis EMG selepas rawatan menunjukkan bahawa berat badan dibahagian kaki belakang dialihkan kembali ke kaki kanan; kembali kepada nilai-nilai pra-OA pada kedua-dua kumpulan rawatan sel (p <0.05). Ciri-ciri mekanikal rawan yang dihasilkan dalam kajian ini dan diambil daripada kawasan yang berbeza adalah setanding dengan rawan asli berdasarkan modulus mampatan dan kekuatan. Tisu rawan yang dihasilkan dalam kajian ini memiliki bukti-bukti korelasi fungsian dengan regenerasi struktur mereka. Kesimpulannya, sel autologous ADSC dan BMSC yang teraruh mempunyai potensi dalam pertumbuhan semula rawan pada OA.,Ph.D. | |
| dc.language.iso | eng | |
| dc.publisher | UKM, Kuala Lumpur | |
| dc.relation | Faculty of Medicine / Fakulti Perubatan | |
| dc.rights | UKM | |
| dc.subject | Osteoarthritis | |
| dc.subject | Adipose stem cells | |
| dc.subject | Bone marrow stem cells | |
| dc.subject | Dissertations, Academic -- Malaysia | |
| dc.subject | Universiti Kebangsaan Malaysia -- Dissertations | |
| dc.title | Chondrogenic induced Adipose derived stem cells and bone marrow stem cells for the treatment of surgically induced Osteoarthritis in a sheep model | |
| dc.type | Theses | |
| dc.format.pages | 224 | |
| dc.identifier.callno | QT20.5.C634c 2014 9HUKMPRA tesis | |
| Appears in Collections: | Faculty of Medicine / Fakulti Perubatan | |
Files in This Item:
| File | Description | Size | Format | |
|---|---|---|---|---|
| ukmvital_80557+SOURCE1+SOURCE1.0.PDF Restricted Access | 54.38 MB | Adobe PDF |  View/Open |
Items in DSpace are protected by copyright, with all rights reserved, unless otherwise indicated.