In vitro and in vivo gene expression studies of cartilage-like tissue engineered construct using a combination of transiently transfected human osteoarthritic chondrocytes and tissue engineering technique

Purpose: To evaluate the formation of cartilage-like tissue engineered constructs (TECs) using human osteoarthritic chondrocytes overexpressed with SOX9 gene seeded on poly(lactic-co-glycolic acid) (PLGA) with and without fibrin scaffolds through cartilaginous genes expression analysis. Methods: Upon approval by the IREC18; NMRR-12-1383-14531 and IIUM/IACUCApproval/2015/[5][22], six cartilage samples were obtained from consented patients after joint replacement surgery. The cells were isolated, cultured and transfected with pcDNA3-SOX9 using Lipofectamine 2000™. Prefabricated disc-shaped porous PLGA with and without fibrin were used as scaffolds. The ‘cells-scaffolds’ TECs were formed and cultured for 3-week and implanted subcutaneously at the dorsum of athymic mice for 4-week. Collagens I, II, IX, X, XI, SOX9 and aggrecan expression were evaluated using a qualitative two-step reverse-transcriptase PCR. GAPDH and β-actin genes were used as internal controls. Results: Presence of cartilaginous markers can be detected in all TECs with various expression intensity. Collagen II, the cartilage-specific marker was down-regulated in vitro but re-expressed in vivo. Collagen I, X, SOX9 and aggrecan were steadily expressed in all TECs. Although collagen IX and XI are closely associated with collagen II, their expressions were almost untraceable except for few cases. Presence of GAPDH and β-actin genes indicated the reliability of the analysis. Conclusion: Cartilage-like TECs have been successfully formed based on genes expression analysis

Total knee replacement: 12 years retrospective review and experience

This retrospective study was undertaken to evaluate the outcome of primary total knee arthroplasty (TKA)performed from January 1986 to December 1997 at this institution. Case review included Knee Society scores and functional knee score. The records of 94 patients (128 knees) were available for analysis with the sample comprised of 76 females (80.9%) and 18 males (19.1%) and a mean age was 61.4 years. TKA was performed for osteoarthritis (OA) in 96 knees (75%) and rheumatoid arthritis (RA) in 32 knees (25%). Cemented TKAs were performed in all patients. The mean knee score improved from 38.8 preoperatively to 90.9 postoperatively. The mean functional score improved from 19.1 preoperatively to 62.5 postoperatively. Both scores showed significant improvement when comparing preoperative to post operative results (p< 0.005). Arthroplasty was designated failures if the prostheses used had been removed. Survivorship at 12 years was 89.5%

Bioartificial Articular Cartilage Substitute (BACS): advancing functional tissue engineered medical product

Articular cartilage has poor repair capacity due to its avascular and aneural properties. To date, there is no definitive treatment that can delay or reverse the degenerative process in cartilage. Therefore this bioartificial articular cartilage substitute is one of the alternatives that employ essential tissue engineering triad, which consist of i) quality cell source i.e. autologous bone marrow mesenchymal stem cells (BMSCs), ii) biomaterial scaffolds i.e. synthetic poly(lactic-co-glycolic)acid (PLGA) [+] natural fibrin and iii) signalling factors (chemical and/or physical stimuli) i.e. selected growth medium. All threecomponents are combined together forming PLGA-based scaffolds and BMSCs to restore damaged joint cartilage. Our implant model demonstrated promising findings with satisfactory structural and functional cartilaginous tissue restoration based on the International Cartilage Repair Society (ICRS) classification and the modified O’Driscoll histological grading scale. This reliable autologous matrix-induced cartilage implantation system uses one’s own cells, therefore the approach can be used to bypass certain social and scientific issues surrounding the conventional tissue or organ transplantation

Midterm outcome of total knee arthroplasty in East Coast population of Hospital Tengku Ampuan Afzan Kuantan

Introduction: Total knee replacement (TKR) is an excellent advancement in medical treatment which gives good joint function and pain relief for degenerative diseases of joint since it was introduced in Malaysia since 1970. The purpose of this study was to evaluate the midterm functional outcome of patients undergone TKR using the NexGen LPS Flex prosthesis between January 2005 until April 2006 in primary osteoarthritis patients in our local east coast population. Materials and Methods: Forty six patients, 87% female (n=40) and 13% male (n=6), aged 54 to 81 (mean, 69±5.69) years underwent primary TKR using the hi-flex knee prosthesis (Zimmer) performed by two different surgeons were followed up for a minimal of five years. All used the same technique of surgery, medial parapatellar incision, cruciate substitute and cemented. Assesment were made based on the quality of life outcome score (SF-36) and the Oxford-12 knee scoring system questionnaires. Results: Oxford-12 revealed that patients had mild problems in terms of function and pain with the average function and pain score of 14.35±4.21 and 9.28±3.01 compared to a maximum possible score of 35 and 25 respectively. For eight health domains of SF-36, patients had the highest score (i.e. good health) for role emotional (RE), followed by mental health (MH), and social functioning (SF). This resulted in a higher score for mental component summary (MCS) than physical component summary (PCS). Since a low score of Oxford-12 indicates a better state of health compared to high score for SF-36, negative correlation between them was expected. Conclusion: The NexGen LPS-Flex TKR had good midterm functional outcome. Overall, TKR were found to be effective in terms of improvement in health-related quality-of-life dimensions

Knee Tuberculosis: a challenge in diagnosis and treatment

Tuberculosis (TB) is an ancient disease dated back from ancient Greece time. Once rare in developed countries, now it has re-emerged due to immigration and secondary immunodeficiency. A 27-year-old lady had left knee pain for the past 4 years, went for knee diagnostic arthroscopy procedure, and diagnosed as pigmented villonodular synovitis (PVNS) of the left knee. Despite regular analgesics and physiotherapy, patient symptoms worsen. Two years after the initial treatment, the patient went for a further workout and diagnosed as knee tuberculosis, commenced on anti-TB treatment, the patient still left complicated with a stiff knee and fixed flexion deformity. Identification of knee tuberculosis during the initial phase is crucial as late diagnosis and treatment will leave the patient with debilitating complications

Kimura’s disease with atypical musculoskeletal presentation

We would like to highlight an unusual clinical presentation of Kimura’s disease, a rare, benign, idiopathic condition, usually seen as swelling and lesions of the head and neck region and commonly affecting young men of Asian descent. It is rare for this disorder to present with lesions on other parts of the body, especially the lower limbs. We report a 27- year-old man who presented with a large mass located at the inguinal region and extending down to the upper thigh. Due to the large size of the mass (28 x 18 cm), our provisional diagnosis was a soft tissue sarcoma. Open biopsy showed that the lesion was benign

Treatment of osteomyelitis in New Zealand white rabbit using impregnated biomaterials with gentamicin: in vivo study outcome of bacterial study and micro-computed topography evaluation

INTRODUCTION: Treatment of osteomyelitis is still a major challenge in orthopaedic field. New Zealand White rabbit is acceptable experimental model described to develop effective local delivery of antibiotics osteomyelitis treatment as it can mimics the disease process in human. The objective of the study is to evaluate the outcome treatment of osteomyelitis in rabbit femur with impregnated Hydroxyappatite Gentamicin. Evaluation at stipulated durations using microCT Skyscan 1176 system. MATERIALS & METHODS: Osteomyelitis is created by inoculation of Staphylococcus aureus ATCC 25923 into rabbit’s distal femur. The rabbit were divided with 3, 6, 12, 26 week assessments. The response of the biomaterials (hydroxyapatite) impregnated with gentamicin against the infection were evaluated through microcomputed topography using Skyscan 1176 system.The bone analysis data were obtained by the software provided by Skyscan 1176 system. The distal femoral shaft were scanned using A1 1mm filter with 18 µm pixel resolutions. Bacterial study of culture and sensitivity, coagulase test, catalase test and gram stain were done to determine the treatment is effective. RESULTS: The data of bone analysis were for tissue volume (TV), bone surface (BS), bone volume (BV), and trabecular thickness (Tb.Th) in all groups were illustrated at Table 1. The bone healing noted in the treated group. The results showed no bacterial were grow in the agar and all test were negative at 26 weeks interval. Table 1 showing the bone analysis through Skyscan1176 system. DISCUSSIONS: Osteomyelitic changes were noted in all rabbit after inoculation of bacteria at week three and six. The micro-computed topographic analysis at 3, 6, 12 and 26 weeks of interval, showed increase bone parameters in treated femur. The bone volume (BV) was increased with value 124.96 mm3 compared to sham group (no treatment given) with bone volume 66.96 mm3. The density of biomaterials was decreased with duration of studies. This data showed that micro-computed topography studies is capable to provide information of bone microstructures in rapid and non-destructive way. The findings showed that impregnated biomaterials with gentamicin have potential to be used in osteomyelitis infection. CONCLUSION: Computed topography results showed at 26 weeks of treatment, the impregnated biomaterials have a potential in treatment of osteomyelitis

Poly(lactic-co-glycolic acid) and atelocollagen hybrid scaffold seeded with annulus fibrosus cells enhances the formation of cartilaginous tissue engineered construct in vitro

Purpose: To evaluate the in vitro formation of 3D tissue engineered constructs (TECs) using rabbits’ annulus fibrosus (AF) cells seeded on poly(lactic-co-glycolic acid) (PLGA) based scaffolds. Methods: Porous disc-shaped PLGA was fabricated using solvent casting and salt leaching technique. It was crosslinked with atelocollagen to form “PA”scaffold group. Fibrin was added to PLGA and PLGA-atelocollagen composite to form “PF” and “PAF” scaffolds, respectively. The AF cells were seeded into the prefabricated scaffolds (1.0x105 cells per scaffold) to form the following TECs groups: AF+PLGA (AFP; control), AF+PLGA+atelocollagen (AFPA), AF+PLGA+fibrin (AFPF) and AF+PLGA+atelocollagen+fibrin (AFPAF). The resulting TECs were cultured for three-week and evaluated for cells viability using MTT assay, cellular morphology and attachment using SEM, cartilaginous matrix production using sGAG assay and DNA content using PicoGreen® assay. Results: Significant number of viable cells was observed in the AFPAF group (987,985.7±286,858.9 cells)when compared to other TECs(AFP: 373,319.0±5,456.9; AFPA: 547,763.4±66,038.2; AFPF: 463,763.4±46,160.8 cells). Cellular morphology and attachment were comparable in all TECs. The AFPA has the highest sGAG accumulation (0.279±0.117mg/ml) but shows no statistical difference when compared to the other TECs (AFP: 0.083±0.038; AFPF: 0.237±0.131; AFPAF: 0.181±0.024 mg/ml).The AFPF has the highest DNA content (1,919.338±89.050 ng/ml) but shows no statistical difference when comparedto the other TECs (AFP: 485.659±27.468; AFPA: 845.987±82.134; AFPAF: 1,575.007±307.174 ng/ml). Hence, atelocollagen seemed to provide better environment for cellular attachment and proliferation. This unique collagenous material also promotes sGAG production and DNA content in TECs. Conclusion: The incorporation of atelocollagen into PLGA scaffold enhances the formation of TECs in vitro

Development of OSH risk assessment in IIUM orthopaedics research laboratory according to ISO/IEC 17025:2017 accreditation

Orthopaedics Research Laboratory accommodate with biocompatibility testing to meet the requirements of ISO/IEC 17025 for medical device. Towards the implementation of the guideline, laboratory safety is the main issue encountered to reduce the risk of hazardous among the laboratory personal This study aimed to ensure the good practices in safety, health and environment are implemented in ORL according to ISO/IEC 17025: 2017 standard guidelines. The hazard identification at ORL starts from the receiving of laboratory animal, New Zealand White Rabbit (NZWR), animal surgery, radiology assessment and euthanasia. There are many dangers found in all these procedures. In this study, only four safety hazards, two health hazards and one environmental hazard was taken and discussed therein by taking risk assessment rating in ORL. Next, the risk control for each danger was identified and recorded. Only four risk controls were highlighted for each hazard encountered by each procedure. The amounts that the average risk level in the procedures studied in ORL are 2.46 which is in tolerate level. However, since the value is close to 3.0, the level of risk is relatively low and is approaching to moderate. The various risk levels for this study are 2.0. The difference between the highest (operating procedures) and the lowest (euthanasia) which showed the difference of non-essential data. There was no significant difference between the hazards found in ORL with all the procedures studied in one-way ANOVA analysis (F:(3,20) = 0.649, p = 0.592, η2p = 0.09). However, risk assessment may change from time to time depending on the hazard detection and it is strongly recommended that some of the stated control measures be taken to reduce future risk levels that could harm to individuals, organizations and countries