Rekayasa Tulang Rawan Sendi (Cartilage Tissue Engineering)

Kerusakan tulang rawan sendi (cartilage) merupakan masalah yang berat karena apabila terjadi kerusakan pada tulang rawan sendi maka tidak akan pernah mengalami kesembuhan. Ini terjadi karena kemampuan regenerasi tulang rawan yang terbatas dan buruk. Regenerasi pada tulang rawan sendi yang buruk terjadi karena sifatnya avaskuler, minimal sel, tanpa membran basal dan tanpa inervasi dari syaraf sehingga nutrisinya hanya tergantung dari proses difusi. Kerusakan cartilage yang menembus subkondral (fullthickness cartilage defect) meskipun terbentuk clot formation karena adanya aliran darah, tetapi tanpa suatu intervenssi akan menghasilkan regenerasi berupa fibrocartilage yang lemah secara biomekanika dan akan mengalami kerusakan setelah beberapa waktu

Regeneration Mechanism of Full Thickness Cartilage Defect Using Combination of Freeze Dried Bovine Cartilage Scaffold - Allogenic Bone Marrow Mesenchymal Stem Cells - Platelet Rich Plasma Composite (SMPC) Implantation

Cartilage defect has become serious problem for orthopaedic surgeon and patients because of its difficult healing that might occur when articular cartilage damage never reach subchondral layer. In this study, we used combination of freeze dried bovine cartilage (FDBC) scaffold, bone marrow mesenchymal stem cells (BM-MSCs), and platelet rich plasma (PRP) composite (SMPC) implanted in full thickness cartilage defect. This study is to explain its regeneration mechanism. This is true experimental research with post-test only control group design using New Zealand White Rabbit. 50 rabbits is divided into three groups of SMPC, BM-MSCs and FDBC. 37 rabbits evaluated after twelve weeks. Histopathologic examination showed the number of chondrocytes, collagen thickness and cartilage width are highest on SMPC group. Immunohistochemical examination showed SMPC group has the highest number of chondroprogenitor cells express FGF-2R, Sox-9, and MAPK. Brown Forsythe test resulted in significant increase the number of chondrocytes (p=0,010), collagen thickness (p=0,000), and cartilage surface width (p=0,015), and increase FGF-2R (p=0,000), MAPK (p=0,000), and Sox-9 (p=0,000) on SMPC group. Using path analysis, there is strong influence from FGF-2R, MAPK, and Sox-9 to the increase of chondrocytes, collagen thickness, and cartilage surface width. Hence, SMPC implantation mechanism of full thickness cartilage defect regeneration can be explained

Efek Ekstrak Kulit Manggis (Garcinia mangostana L.) Pada Ekspresi Dari TLR5 Dan CD14 Pada Mencit Yang Diberi Vaksin Newcastle Disease

Mangosteen pericarp has well-recognized for reducing the incidence of degenerative diseases including cancer, heart disease, inflammation, arthritis, and immune system. The main chemical substance of mangosteen pericarp that role in improving health is the xanthone which is belonged to phenolic acid that has been studied for its remarkable biological activities in recent years. The research was conducted to analyze the immunomodulatory effect of mangosteen pericarp extract (MPE), by measuring both the expression of TLR5 and CD14 in mice PBMCs. Animal used in the research were 36 female Balb/c mice, that randomly separated into six groups (T0, T1, T2, T3, T4, T5) with six animal each. T0 was negative control group, TI was administered with 40 mg/ml MPE, T2 until T4 was administered with 20 mgml, 40 mg/ml, 60 mg/ml respectively, and T5 was positive control group using Stimuno®. All groups were vaccinated by inactive velogenic type ND vaccine except for T1 group once and without booster. Blood collection has been done a week after vaccination. The result indicated that MPE could increase both the activity of TLR5 and CD14 with 40 mg/ml as optimal dose Keywords: mangosteen pericarp extract, immunomodulatory effect, mice, TLR5, CD1

The Osteogenic Capacity of Human Amniotic Membrane Mesenchymal Stem Cell (hAMSC) and Potential for Application in Maxillofacial Bone Reconstruction in Vitro Study

Amniotic membrane of human placenta is a source of abundant mesenchymal stem cell (hAMSC) which makes it a potential source of allogeneic multipotent cell for bone healing. However, much has to be explored about its isolation procedure and the osteogenic differentiation potential. The aims of this study are to establish the procurement procedure of human amniotic membrane, the isolation and culture of hAMSC, the MSC phenotypic characterization, and the in vitro osteogenic differentiation of hAMSC. Results of the study are as follows. The quality of human amniotic membrane would be best if procured from Caesarean operation under highly aseptic condition to avoid fungal and bacterial contamination on the culture. Isolation procedure using modified Soncini protocol yielded large amount of MSC with high proliferative capacity in culture medium. Characterization of hAMSC showed that the majority of the target cells exhibited specific MSC markers (CD105 and CD90) with a small number of these cells expressing CD45, the marker of hematopoeitic cells. The in vitro osteogenic differentiation of hAMSC followed by Alizarin Red staining showed that osteoblastic differentiation was detected in a significantly high number of cells. This study concludes that hAMSCs isolated from human amniotic membrane have the capacity for in vitro osteogenesis which makes them be one of the potential allogeneic stem cells for application in maxillofacial bone reconstruction

Regeneration Mechanism of Full Thickness Cartilage Defect Using Combination of Freeze Dried Bovine Cartilage Scaffold - Allogenic Bone Marrow Mesenchymal Stem Cells - Platelet Rich Plasma Composite (SMPC) Implantation

Cartilage defect has become serious problem for orthopaedic surgeon and patients because of its difficult healing that might occur when articular cartilage damage never reach subchondral layer. In this study, we used combination of freeze dried bovine cartilage (FDBC) scaffold, bone marrow mesenchymal stem cells (BM-MSCs), and platelet rich plasma (PRP) composite (SMPC) implanted in full thickness cartilage defect. This study is to explain its regeneration mechanism. This is true experimental research with post-test only control group design using New Zealand White Rabbit. 50 rabbits is divided into three groups of SMPC, BM-MSCs and FDBC. 37 rabbits evaluated after twelve weeks. Histopathologic examination showed the number of chondrocytes, collagen thickness and cartilage width are highest on SMPC group. Immunohistochemical examination showed SMPC group has the highest number of chondroprogenitor cells express FGF-2R, Sox-9, and MAPK. Brown Forsythe test resulted in significant increase the number of chondrocytes (p=0,010), collagen thickness (p=0,000), and cartilage surface width (p=0,015), and increase FGF-2R (p=0,000), MAPK (p=0,000), and Sox-9 (p=0,000) on SMPC group. Using path analysis, there is strong influence from FGF-2R, MAPK, and Sox-9 to the increase of chondrocytes, collagen thickness, and cartilage surface width. Hence, SMPC implantation mechanism of full thickness cartilage defect regeneration can be explained

Healing Mechanism and Osteogenic Capacity of Bovine Bone Mineral—Human Amniotic Mesenchymal Stem Celland Autogenous Bone Graft in Critical Size Mandibular Defect

Experiments on maxillofacial bone tissue engineering showed the promising result; however, its healing mechanisms and effectiveness had not been fully understood. The aim of this study is to compare the bone healing mechanism and osteogenic capacity between bovine bone mineral loaded with hAMSC and autogenous bone graft in the reconstruction of critical size mandibular bone defect. Critical size defects were made at the mandible of 45 New Zealand white rabbits reconstructed with BBM-hAMSC, BBM alone, and ABG, respectively. At the end of first, second, and twelfth weeks, five rabbits from each experimental week were sacrificed for histology and immunohistochemistry staining. Expressions of vascular endothelial growth factor (VEGF), bone mor-phogenic proteins-2 (BMP2), Runx2 and the amount of angiogenesis were analyzed in the first and second week groups, while expressions of Runx2, osteocalcin, collagen type-I fibres, trabecular area and bone incorporation were analyzed in the twelfth week groups. The result showed that expressions of VEGF, BMP2 and Runx2 as well as the amount of angiogenesis were higher in ABG compared with BBM-hAMSC group in the first and second weeks of healing. The result of twelfth week of healing showed that expressions of Runx2 and osteocalcin as well as the thickness of collagen type-I fibres were significantly higher in BBM-hAMSC compared to ABG group, while there was no statistically difference in trabecular area and bone incorporation between BBM-hAMSC and ABG group. This study concluded that early healing activities were higher in auto-genous bone graft than in BBM-hAMSC, while osteogenic activities in the late stage of healing were higher in BBM-hAMSC compared to autogenous bone graft. It was also concluded that the osteo-genic capacity of BBM-hAMSC was comparable to autogenous bone graft in the reconstruction of critical size defect in the mandible

EKSPRESI PROTEIN P53 DAN HSP70 PADA SEL PUNCA KARSINOMA NASOFARING YANG RESISTEN TERHADAP RADIOTERAPI

Latar belakang: Pada penderita Karsinoma Nasofaring (KNF) masih sering ditemukan kekambuhan meskipun sudah mendapat terapi yang lengkap. Penelitian terbaru membuktikan bahwa kekambuhan disebabkan oleh sel punca KNF yang resisten terhadap radioterapi. Mekanisme resistensi sel punca kanker terhadap radioterapi diduga karena hambatan terhadap apoptosis dan atau memicu proliferasi. Hambatan terhadap apoptosis disebabkan oleh penurunan protein p53 (wild type), selain over-ekspresi Hsp70. Tujuan: Menjelaskan mekanisme resistensi sel punca KNF terhadap radioterapi berdasarkan profil ekspresi protein p53(wild type)dan Hsp70. Metode: Penelitian true experimental dengan menggunakan rancangan randomisasi kelompok kontrol sebelum dan sesudah tes. Kultur sel punca KNF dibagi menjadi dua kelompok, masing-masing 16 sampel. Pada kelompok perlakuan diberikan paparan radioterapi dengan dosis 1,5 Gy menggunakan pesawat Linac, lalu diinkubasi selama 24 jam. Sebelum dan sesudah perlakuan pada kedua kelompok diperiksa ekspresi p53 (wild type) dan Hsp70. Pemeriksaan menggunakan metode flowcytometry. Hasil: Ekspresi p53 (wild type) antara kelompok perlakuan dan kontrol terdapat perbedaan yang tidak bermakna dengan p=0,576 (p≥0,05). Ekspresi Hsp70, antara kelompok perlakuan dan kontrol terdapat perbedaan yang tidak bermakna dengan p=0,172 (p≥0,05). Kesimpulan: Tidak terdapat perubahan ekspresi p53 (wild type) dan Hsp70 pada sel punca KNF yang resisten terhadap radioterapi

Foam-Cell Signified Blood Vessel Endhotel Repair and Histopatology of Abdominal Aorta through Stem Cell Allogenous Therapy to Rats (Rattus norvegicus) with Atherosclerosis

Atherosclerosis is a chronic inflammation process of endothel cell layer of blood vessels which is initiated by the disfunction of the endothel. This research aimed at understanding the repairment mechanism of the function of endothel in cardiac blood vessels with ateroskleroris case after being given medium-intenity physical exercises, mesenchymal stem cell and combination of the medium intensity physical exercises and mesenchymal stem cell by lookin into the foam cell of abdominal aorta. This research employed true experimental research design with post test only control group design. The sample of this reseach were 24 male Wistar rats (Rattus norvegicus) furrow that were controlled its homogeneity using inclusive criteria; confirming ateroclerosis, 20 week age, weight ranged from 180-200 gram, inhybrid, and healthy that were indicated by good desire for food and behaved normally. The Rattus norvegicus which fulfilled the inclusive criteria were divided into three groups which first group was the control group (atheroscleoris rats). The second group was ateroclerosis rats and received regular medium-intensity physical exercises. The third group atherosclerosis which received combination of regular medium-intensity physical exercises and received mesenchymal stem cell. The result of manova test showed value p < 0.001 which indicated the existence of different foam cell found in the control group, exercise group, stem cell group and combined exercise and stem cell group. It can be concluded that attempt to decrease the risk factor of aterosclerosis is one of the ways to protect the endothel of the blood vessels. Deep understanding on this mechanism is expected to give new insights to do preventive action and treatments toward ateroclerosis by combination theraphy of regular medium-intensity physical exercises and received mesenchymal stem cell

Human Umbilical Cord Mesenchymal Stem-Cell Therapy to Increase the Density of Osteoporotic Mandibular Bone

Objective The aim of this study is to evaluate the feasibility of human umbilical cord mesenchymal stem-cell (hUCMSC) therapy in increasing osteoporotic mandibular bone density in a rat model by determining changes in alkaline phosphatase (ALP), osteocalcin, type 1 collagen, and trabecular bone area after treatment. Materials and Methods This research adopted an experimental posttest-only control group design. Thirty female Wistar rats were randomly divided into six groups, namely, a control group with rats postsham surgery (T1), osteoporotic model postovariectomy rats (T2), postovariectomy rats 4 weeks after gelatin injection (T3), postovariectomy rats 8 weeks after gelatin injection (T4), postovariectomy rats 4 weeks after hUCMSC injection (T5), and postovariectomy rats 8 weeks after hUCMSC injection (T6). The rats were all sacrificed for histological and immunohistochemical examinations of ALP, osteocalcin, type 1 collagen, and trabecular bone area. Results Increased expression of ALP, type 1 collagen, and osteocalcin, as well as increased trabecular bone area, was observed in the treatment groups compared with that in the osteoporotic groups. Conclusion hUCMSCs produce significant osteogenic effects and increase osteoporotic mandibular bone density in the animal model. Increases in bone density are demonstrated by the higher levels of ALP, osteocalcin, and type 1 collagen, as well as increases in the trabecular bone area

Early Graft Tunnel Healing After Anterior Cruciate Ligament Reconstruction With Intratunnel Injection of Bone Marrow Mesenchymal Stem Cells and Vascular Endothelial Growth Factor

Background: Bone marrow mesenchymal stem cells (BM-MSCs) are multipotent adult stem cells and have become an important source of cells for engineering tissue repair and cell therapy. Vascular endothelial growth factor (VEGF) promotes angiogenesis and contributes fibrous integration between tendon and bone during the early postoperative stage. Both MSCs and VEGF can stimulate cell proliferation, differentiation, and matrix deposition by enhancing angiogenesis and osteogenesis of the graft in the tunnel. Hypothesis: Injection of intratunnel BM-MSCs and VEGF enhances the early healing process of a tendon graft. Study Design: Controlled laboratory study. Methods: In this controlled animal laboratory study, each of 4 groups of rabbits underwent unilateral anterior cruciate ligament (ACL) reconstruction with use of the ipsilateral semitendinosus tendon. The rabbits received intratunnel injection of BM-MSCs and VEGF with a fibrin glue seal covering the distal tunnel at the articular site. Evaluation using magnetic resonance imaging (MRI), collagen type III expression, and biomechanical analyses were performed at 3- and 6-week intervals. Results: All parameters using MRI, collagen type III expression, and biomechanical analysis of pullout strength of the graft showed that application of intratunnel BM-MSCs and VEGF enhanced tendon-to-bone healing after ACL reconstruction. Conclusion: Intratunnel injections of BM-MSCs and VEGF after ACL reconstruction enhanced graft tunnel healing. Overall, the femoral tunnel that received BM-MSCs and VEGF had better advanced healing with increased collagen type III fibers and better outcomes on MRI and biomechanical analysis. MRI is the most reliable tool for clinical use in evaluating stages of ACL healing after reconstruction, since biopsy is an invasive procedure