Recent advancement in cartilage tissue engineering has explored the potential of 3D culture to mimic the in vivo environment of human cartilaginous tissue. Three-dimensional culture using microspheres was described to play a role in driving the differentiation of mesenchymal stem cells to chondrocyte lineage. However, factors such as mechanical agitation on cell chondrogenesis during culture on the microspheres has yet to be elucidated. In this study, we compared the 2D and 3D culture of bone-marrow-derived mesenchymal stem cells (BMSCs) on gelatin microspheres (GMs) in terms of MSC stemness properties, immune-phenotype, multilineage differentiation properties, and proliferation rate. Then, to study the effect of mechanical agitation on chondrogenic differentiation in 3D culture, we cultured BMSCs on GM (BMSCs-GM) in either static or dynamic bioreactor system with two different mediums, i.e., F12: DMEM (1:1) + 10% FBS (FD) and chondrogenic induction medium (CIM). Our results show that BMSCs attached to the GM surface and remained viable in 3D culture. BMSCs-GM proliferated faster and displayed higher stemness properties than BMSCs on a tissue culture plate (BMSCs-TCP). GMs also enhanced the efficiency of in-vitro chondrogenesis of BMSCs, especially in a dynamic culture with higher cell proliferation, RNA expression, and protein expression compared to that in a static culture. To conclude, our results indicate that the 3D culture of BMSCs on gelatin microsphere was superior to 2D culture on a standard tissue culture plate. Furthermore, culturing BMSCs on GM in dynamic culture conditions enhanced their chondrogenic differentiation.

Long urethral strictures are often treated with autologous genital skin and buccal mucosa grafts; however, risk of hair ingrowth and donor site morbidity, restrict their application. To overcome this, we introduced a tissue-engineered human urethra comprising adipose-derived stem cell (ASC)-based self-assembled scaffold, human urothelial cells (UCs) and smooth muscle cells (SMCs). ASCs were cultured with ascorbic acid to stimulate extracellular matrix (ECM) production. The scaffold (ECM) was stained with collagen type-I antibody and the thickness was measured under a confocal microscope. Results showed that the thickest scaffold (28.06 ± 0.59 μm) was achieved with 3 × 104 cells/cm2 seeding density, 100 μg/mL ascorbic acid concentration under hypoxic and dynamic culture condition. The biocompatibility assessment showed that UCs and SMCs seeded on the scaffold could proliferate and maintain the expression of their markers (CK7, CK20, UPIa, and UPII) and (α-SMA, MHC and Smootheline), respectively, after 14 days of in vitro culture. ECM gene expression analysis showed that the ASC and dermal fibroblast-based scaffolds (control) were comparable. The ASC-based scaffold can be handled and removed from the plate. This suggests that multiple layers of scaffold can be stacked to form the urothelium (seeded with UCs), submucosal layer (ASCs only), and smooth muscle layer (seeded with SMCs) and has the potential to be developed into a fully functional human urethra for urethral reconstructive surgeries.

Traditionally, foetal bovine serum (FBS) is used as a serum supplement for stem cell expansion in vitro. However, it is associated with xenoimmunisation and the transmission of animal pathogens, which may cause harm to stem cell recipients. As a safer alternative, human platelet lysate (HPL) has been introduced for propagating stem cells. Chondrocytes are expanded in vitro for cartilage repair via autologous chondrocyte implantation (ACI). In this study, we compare the efficacy of HPL prepared from expired platelet concentrates with that of FBS for promoting the proliferation and maintenance of the chondrogenic markers of primary human chondrocytes expanded in vitro. Chondrocytes were cultured in F12:Dulbecco's modified Eagle's medium (DMEM) supplemented with 5% HPL, 10% HPL and 10% FBS. The cell morphology, viability and growth rate were examined from passage 1 (P1) to P3. RNA was isolated from P3 cells for quantitative polymerase chain reaction (qPCR) to determine the gene expression level of the chondrogenic, dedifferentiation and hypertrophic markers. HPL promoted chondrocyte proliferation without compromising cell viability. In addition, the chondrocytes cultured with HPL were smaller. However, HPL failed to maintain the chondrogenic markers, except SOX 9 (SRY-box transcription factor 9), which was upregulated, but not significantly. Nonetheless, HPL also suppressed the expression of type X collagen (Col X), a chondrocyte hypertrophic marker. In summary, we demonstrate the benefits of HPL supplementation in human chondrocyte culture, where it enhances cell proliferation and suppresses chondrocyte hypertrophy. In the future, HPL can be used for the large-scale expansion of chondrocytes for ACI. ABSTRAKSecara tradisinya, serum anak lembu (FBS) digunakan untuk pengkulturan sel in vitro. Namun demikian, FBS dikaitkan dengan penolakan oleh sistem imun dan penularan patogen haiwan yang berkemungkinan membawa kemudaratan kepada penerima sel induk. Lisat platelet manusia (HPL) telah diperkenalkan sebagai alternatif yang lebih selamat untuk pengkulturan sel. Sel kondrosit telah dikultur secara in vitro untuk merawat rawan yang rosak melalui teknik implantasi kondrosit autologus (ACI). Pada kajian ini, kami membandingkan keberkesanan HPL yang disediakan daripada platelet pekat tamat tempoh dan FBS dalam merangsang proliferasi dan mengekalkan penanda kondrogenik sel kondrosit manusia yang dikulturkan secara in vitro. Sel kondrosit dikultur dalam medium F12:DMEM yang ditambah dengan 5% HPL, 10% HPL dan 10% FBS. Morfologi, kadar kehidupan dan kadar pertumbuhan sel daripada P1 hingga P3 dikenal pasti. RNA dipencil daripada sel pada P3 untuk qPCR untuk menentukan ekspresi gen penanda-penanda kondrogenik, penyahbezaan dan hipertrofi. Hasil kajian mendapati HPL merangsang proliferasi sel kondrosit tanpa memkompromikan kadar kehidupan sel. Selain itu, sel kondrosit yang dikultur dengan HPL menunjukkan saiz yang lebih kecil. Namun demikian, HPL gagal mengekalkan penanda-penanda kondrogenik kecuali SOX 9 yang meningkat secara t...

Total Knee Replacement (TKR) is a treatment of knee osteoarthritis to alleviate pain symptoms and improve physical functioning when other conservative treatments have failed. However, some patients still complaining of persistent post surgical pain (PPSP) after TKR. Although the cause of PPSP is not known, it seems that some of the patients with PPSP after TKR suffer from neuropathic pain. Recently genicular nerve radio-frequency ablation has emerged as an alternative intervention for patients at high risk who are unable to undergo surgical intervention. We have shown from our case, a 69-year old lady who had right total knee replacement presented with neuropathic knee pain has good symptom relieves after undergone radio-frequency ablation.

Introduction: To evaluate the midterm functional outcomes of patients who had undergone total knee replacement (TKR) using posterior stabilized (PS) fixed bearing system from January 2005 until April 2006 for primary osteoarthritis of the knees. Methods: A retrospective study was done on forty-six patients, 87% female (n= 40) and 13% male (n=6), aged 54 to 81 years old (mean 69 ± 5.69) who had undergone primary TKR using the NexGen LPS-Flex Zimmer Warsaw. The patients were followed up for at least five years. A medial parapatellar approach and cemented posterior stabilized fixed bearing TKR system were used performed by one of two arthroplasty surgeons. All patients followed the same rehabilitation protocol post-operatively. Assessments were done using Oxford-12 knee scoring questionnaire to evaluate knee function and SF-36 to assess quality of life outcomes. Results: Oxford knee score revealed patients had lower (better) than average scores in terms of function (score 14.35 ± 4.21 out of 35) and pain (score 9.28 ± 3.01 out of 25). Analysis on the quality of life outcome score with SF-36 revealed a higher (better) score for mental component summary (MCS) relative to physical component summary (PCS). Patients had the highest score for role emotional (RE), followed by mental health (MH), and social functioning (SF). Conclusions: The NexGen LPS-Flex TKR produced a good functional outcome with minimal pain in the operated knee with better mental compared to physical health of patients in the midterm period post-total knee replacement.