Purpose:To investigate the feasibilty of using processed human amniotic membrane (HAM) to support the attachment and proliferation of chondrocytes in vitro which it turn can be utilised as a cell delivery vehicle in tissue engineering applications. Methods: Fresh HAM obtained from patients undergoing routine elective ceasarean sections was harvested., processed and dried using either freez drying (FD) or air drying (AD) methods prior to sterilisation by gamma irradiation. Isolated, processed and characterised rabbit autologous chondrolytes were seeded on processsed HAM and cultured for up to three weeks. Cell attachment and proliferation were examined qualitatively using inverted brightfield microcospy. Results: Processed HAM appeared to allow cell attachment when implanted with chrondocytes. Although cells seeded on AD and FD HAM did not appear to attach as strongly as those seeded on glycerol preserved intact human amniotic membrane, these cells to be proliferated in cell culture conditions. Conclusion: Prelimanary results show that processed HAM chondrocyte attachment and proliferation.
Valvular dysfunction as the prominent reason of heart failure may causes morbidity and mortality around the world. The inability of human body to regenerate the defected heart valves necessitates the development of the artificial prosthesis to be replaced. Besides, the lack of capacity to grow, repair or remodel of an artificial valves and biological difficulty such as infection or inflammation make the development of tissue engineering heart valve (TEHV) concept. This research presented the use of compound of poly-l-lactic acid (PLLA), thermoplastic polyurethane (TPU) and maghemite nanoparticle (γ-Fe₂O₃) as the potential biomaterials to develop three-dimensional (3D) aortic heart valve scaffold. Electrospinning was used for fabricating the 3D scaffold. The steepest ascent followed by the response surface methodology was used to optimize the electrospinning parameters involved in terms of elastic modulus. The structural and porosity properties of fabricated scaffold were characterized using FE-SEM and liquid displacement technique, respectively. The 3D scaffold was then seeded with aortic smooth muscle cells (AOSMCs) and biological behavior in terms of cell attachment and proliferation during 34 days of incubation was characterized using MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assay and confocal laser microscopy. Furthermore, the mechanical properties in terms of elastic modulus and stiffness were investigated after cell seeding through macro-indentation test. The analysis indicated the formation of ultrafine quality of nanofibers with diameter distribution of 178 ± 45 nm and 90.72% porosity. In terms of cell proliferation, the results exhibited desirable proliferation (109.32 ± 3.22% compared to the control) of cells over the 3D scaffold in 34 days of incubation. The elastic modulus and stiffness index after cell seeding were founded to be 22.78 ± 2.12 MPa and 1490.9 ± 12 Nmm², respectively. Overall, the fabricated 3D scaffold exhibits desirable structural, biological and mechanical properties and has the potential to be used in vivo.
Uninfected agarwood branch is readily available as raw material in agarwood plantation as new practices of agarwood plantation scheme were opted as substitute to the endangered wild type agarwood. The uninfected branch can be easily obtained during pruning process (one of plantation’s common maintenance procedure), throughout the years before inoculation stage. This current study aimed to investigate the optimal extraction process conditions of agarwood branch using ethanol as solvent system for maximal yield, and assess its cytotoxic effects towards MCF-7 breast cancer cells. Uninfected branch of Aquilaria subintegra was subjected to One Factor at a Time (OFAT) and Response Surface Methodology (RSM)-guided ethanolic extraction to achieve maximal yield. The extract was then subjected to cytotoxicity, cell attachment and cell viability assays, respectively. Optimization Run 2 (temperature 45 °C, solid-liquid ratio of 1:30, 16 hours maceration) gave the highest agarwood branch ethanolic extract (ABEE) yield of 44.70 ± 18.9 mg/g dried material (DM). Meanwhile Run 7 (temperature 45 °C, solid-liquid ratio of 1:10, 16 hours of maceration) gave the lowest yield (19.34 ± 14.1 mg/g DM). However, while maintaining the 16 hour-maceration, the model predicted a slightly lower yield of 30.232 ± 0.266 mg/g DM of ABEE with process conditions of 45 °C and solid-liquid ratio of 1:19 when the desirable parameters were factored in namely using (ⅰ) the most suitable temperature (that does not risk the bioactivities of the extract), and (ⅱ) an economical volume of solvent. Crude ABEE obtained from the optimal process conditions resulted in cytotoxicity effects on MCF-7 breast cancer cells with IC50 estimate of 3.645 ± 0.099 μg/mL. The extract also affected MCF-7 cell attachment and viability with altered morphology. More work to elucidate the mechanism of actions of the extract are warranted; which could further lead to development of breast cancer natural product-based therapeutics.