Journal of Biomimetics, Biomaterials and Biomedical Engineering Vol. 52

Abstract: Bone tissue is a calcium deposit and supporting structure of the human body, it is exposed to several pathologies that modify its mineral content. To determine these changes, different diagnostic procedures are performed with techniques using invasive ionizing radiation, which are limited by the negative effects in the long term on human health. A methodology is explored that could be applicable in the diagnosis of pathologic variations in bone mineral density, using structural monitoring tools. The proposed technique estimates changes in bone conditions by applying impedance spectroscopy with a tooth-borne piezo-device. Bone-tooth samples were prepared to simulate a section of maxillary bone and subsequently treated with chemical agents, simulating pathologic decalcification. The piezo-device is inserted in the slot of an orthodontic bracket, previously bonded to the crown of the tooth, in order to transmit vibration to surrounding bone. The variations in bone micro-architecture were computed by image processing analyzed with samples prepared in transparent resin, allowing the measurement of morphometry before and after the induced changes in mineral content. Using vibrational bone response, impedance measurements allowed to observe the variations in bone mass as the samples were progressively decalcified. In the 5-50kHz spectrum, it was demonstrated the sensitivity of the electro-mechanical impedance during the bone alteration procedure since the electrical resistance signals of the piezo-device consistently changed in the frequency spectrum (5-50kHz). The piezo-device shows to be sensitive to the changes produced by the bone alterations, which were caused by the stiffness variations made in the sample during the decalcifying. These changes were statistically correlated to demonstrate that in a less invasive way, bone alterations could be monitored from the teeth. This result opens the door to search for a new way to diagnose bone density changes in real applications.

Abstract: Abstract. Zinc oxide nanoparticles (ZnO NPs) are known to be one of the multifunctional inorganic nanoparticles with its application in the treatment of bacterial pathogens, especially when synthesized through green nanotechnology. In this study, ZnO NPs were successfully synthesized through co-precipitation method and its antibacterial activity against Ralstonia solanacearum was evaluated. Surface morphology through scanning electron microscope (SEM) exhibited an agglomerated rod-like structures, with a mean particle size of 180.9 nm. Phytochemical screening was performed through various chemical qualitative tests, to which the presence of terpenoids and cardiac glycosides in Caesalpinia sappan leaves was confirmed in the aqueous extract. Five treatments were evaluated against R. Solanacearum in terms of their zone of inhibition. The highest zone of inhibition from the different concentrations was observed from the positive control (Gentamicin) with a mean value of 34.47 mm, followed by 0.57 g/mL ZnO NPs with a mean value of 21.69 mm, and no zone of inhibition on the negative control, 0.28 g/mL, and 0.19 g/mL of synthesized ZnO NPs. Antibacterial activity of ZnO using disc diffusion method resulted in a significant zone of inhibition which proves that synthesized nanoparticles can be used as a potent antibacterial agent against R. solanacearum.

Abstract: Titanium based metallic biomaterials for orthopedic implant applications are often associated with biocompatibility problems which can be ameliorated via proper surface modification strategies. Improving the hydrophilic nature of the titanium surface offers an effective strategy to sort out such limitations by intensifying the cellular activity. Development of titania as well as titanate layers on the titanium surface via alkali treatment represents an effective strategy to improve the hydrophilicity of native titanium surface. Inspired from nature, in the present work, we report the formation of three-dimensional (3D) hierarchical nanoflowers resembling Gomphrena globosa flowers developed on commercially pure titanium (cp-Ti) surface via a facile alkali treatment technique. X-ray diffraction studies evidenced anatase and rutile phases of TiO₂ confirming the development of titania on the surface. In addition to the TiO₂ phase, presence of titanate (Na₂Ti₃O₇) has also been observed as alkali treatment was conducted in NaOH solution. The hydrophilicity of the Ti surface has been enhanced after the alkali treatment as evidenced from wettability studies using static contact angle measurements. This increase in hydrophilicity is due to the enrichment of the surface by TiO₂ and titanate and increased roughness of nanoflower surface based on classical Wenzel law. In addition, the alkali-treated surface demonstrated an increased polar surface energy beneficial for biocompatible surfaces.

Abstract: Spinal cord injury is damage to the spinal cord which causes lesions in the spinal cord and leads to an increase in extracellular Ca²⁺. It results in additional neuronal loss which causes temporary/permanent disability or even death. The aim of this study was to determine characteristics and the best composition of alginate – chitosan hydrogel responsive to Calcium (Ca²⁺) for spinal cord injury. Hydrogel synthesis with its compositions, namely chitosan was dissolved in 0.4% acetic acid, neutralized in pH 7 with 0.5 M NaOH, added some 0.85% NaCl in it, and added 5 alginate variations which were dissolved in 0.85% NaCl, next will centrifugation method. Based on the FTIR test, hydrogel showed stretching vibrations of Chitosan’s O–H bonds appeared in 3415.93cm^-1 wavenumber, while Na groups of alginate isomer appeared in 1413.82 cm^-1 wavenumber. The results of the cytotoxicity test using the MTT Assay method showed live cell percentage from less than 50% to 52.61% in Sample B and 83.83% in Sample C. The results of the injectability test showed that all samples were injectable with the highest percentage of injectability at 98.283%. The results of the UV-Vis spectrophotometric test showed that all hydrogel samples were able to absorb Ca²⁺. Hydrogels can be degraded at more than 90% within 14 days. The results of the morphology test (SEM) obtained 84.7-99.6 μm pore sizes.

Abstract: Liver biological scaffold was developed in order to resemble native liver tissue environment. It can be achieved by decellularizing native liver tissue that will remove cells and preserve extracellular matrix (ECM). Furthermore, ECM fibers are arranged in a special pattern, which affect liver cell polarity and topography that are important for cells’ implantation, proliferation and differentiation. Therefore, the aim of this study was to evaluate liver cube scaffold topography that was decellularized with fixed multiple sites syringe injection (Indonesia patent number: S00201907930).Rat liver cubes (n=3) underwent decellularization with Ethylene Glycol Tetraacetic Acid (EGTA) immersion and increased Sodium Dodecyl Sulfate (SDS) concentrations using previous multiple sites syringe injection protocol study. Deoxyribonucleic Acid (DNA) concentrations were measured to confirm less DNA materials remaining in scaffolds. Scanning Electron Microscope (SEM) analysis of scaffolds were conducted for topographic characterization compared to undecellularized liver control. Molecular analysis of DNA concentration showed complete removal of DNA material. SEM analysis gave appearance of intact liver cube scaffold microarchitecture. Liver cubes decellularization using multiple sites syringe injection showed good topographic liver scaffold characterization.

Abstract: Intraperitoneal adhesion is a serious case that often occurs with a prevalence of 90-97 % after undergoing gynecological surgery and laparotomy. This study aims are to characterized the hydrogel and identified the optimal composition of Hyaluronic acid (HA) - N, O-carboxymethyl chitosan (NOCC) as an anti-adhesion biomaterial barrier. The synthesis method involved firstly the synthesis of aldehyde derivative of hyaluronic acid (AHA) and also the conversion of chitosan into its derivative, N,O-carboxymethyl chitosan. These two compounds were mixed in various compositions and crosslinked to form N, O-carboxymethyl chitosan (NOCC) /AHA. Fourier-transform infrared spectroscopy has confirmed that the functional groups found -C = O stretching at 1644 cm^-1 indicating the hyaluronic acid and carboxymethyl group (-CH2COOH) in 1380 cm^-1 which indicate the presence of chitosan. The crosslink is evidenced by the group C = N stretching at a wavenumber of about 1630 cm^-1. The best composition of intraperitoneal anti-adhesion is the ratio of hyaluronic acid: chitosan at 30:10 mg/ml. The swelling test is showed a swelling ratio of around 211.8 % in accordance with the standard as intraperitoneal anti-adhesion. Hydrogel has a degradation rate up to 86.87 % on day 10, and this is in accordance with the standard as intraperitoneal anti-adhesion. Cytotoxicity assay showed that hydrogel was nontoxic with a percentage of 92.9 % cell viability. The newly developed hyaluronic acid-carboxymethyl chitosan has characteristics that conform to the criteria of an intraperitoneal anti-adhesion.

Abstract: Titanium (Ti) has been used in metallic implants since the 1950s due to various biocompatible and mechanical properties. However, due to its high Young’s modulus, it has been modified over the years in order to produce a better biomaterial. Tantalum (Ta) has recently emerged as a new potential biomaterial for bone and dental implants. It has been reported to have better corrosion resistance and osteo-regenerative properties as compared to Ti alloys which are most widely used in the bone-implant industry. Currently, Tantalum cannot be widely used yet due to its limited availability, high melting point, and high-cost production. This review paper discusses various manufacturing methods of Tantalum alloys, including conventional and additive manufacturing and also discusses their drawbacks and shortcomings. Recent research includes surface modification of various metals using Tantalum coatings in order to combine bulk material properties of different materials and the porous surface properties of Tantalum. Design modification also plays a crucial role in controlling bulk properties. The porous design does provide a lower density, wider surface area, and more immense specific strength. In addition to improved mechanical properties, a porous design could also escalate the material's biological and permeability properties. With current advancement in additive manufacturing technology, difficulties in processing Tantalum could be resolved. Therefore, Tantalum should be considered as a serious candidate material for future bone and dental implants.

Abstract: MicroRNAs (miRNAs) have ability to down-regulate gene expressions. hsa-miR-17-5p, has been confirmed as an oncogene or tumor suppressor. However, the existence on human adipose-derived stem cells (ADSCs) or adipocytes, is still unclear. Many researchers emphasizing the role of hsa-miR-17-5p on cellular senescence, aging and cancer, but not specific on the expression in the exosome of human ADSCs and adipocytes. The primary ADSCs were derived from subcutaneous adipose tissue of pregnant woman during elective cesarean operation, then processed by combining conventional and enzymatic methods. Adipocytes were differentiated by using the StemPro Adipogenesis Differentiation kit^® and Oil Red-O staining. Exosomes were isolated using Exosome Purification and RNA Isolation kit^® and were characterized by scanning electron microscope. The markers, CD34 and CD44, were identified and analyzed by using FACS analysis. Subsequently, microRNA was extracted and observed for hsa-miR-17-5p expression. This study showed that ADSCs and adipocytes were proved to express CD34⁺ and CD44^+. The hsa-miR-17-5p were also detected in both the exosome of ADSCs and adipocytes. Although the source of the ADSCs was from pregnant woman, the characteristic was similar with the ones from non-pregnant woman. Our study also supports the questionable existence of CD34 in ADSCs. Having confirmed the characteristics, we proved that the exosomes of ADSCs and adipocytes expressed similar hsa-miR-17-5p despite they are from phenotypically different cell types and may have distinct roles. However, further research steps should be done in the future to verify the role of hsa-miR-17-5p towards senescent cell and ADSC differentiation.

Abstract: The use of mesenchymal stem cells can add local improvements potential to enthesis tissue regeneration based on tropical activity through secretions of growth factors, cytokines, and vesicles (e.g. exosomes), collectively known as secretomes. This study aims to analyze secretomes characterization from adipose-derived mesenchymal stem cells seeded with enthesis tissue scaffold in hypoxic conditions and to analyze the influence of hypoxic environment to the characterization of secretomes. This is an in-vitro study using a Randomized Control Group Post-Test Only design. This study using Adipose Stem Cells (ASCs) were cultured in hypoxia (Oxygen 5%) and Normoxia (21%) condition. The scaffolds are fresh-frozen enthesis tissue and was seeded in the treatment group and compared to control. The evaluation of Scleraxis (Scx) and SRY-box (Sox9) was measured using ELISA on the 2nd, 4th, and 6th days. Comparison of Scx levels between each evaluation time showed a positive trend in a group with scaffold in hypoxia condition although it has no significant differences (p=0.085), with the highest level on day 6, that is 13,568 ng/ml. Conversely, the comparison of Sox9 showed significant differences (p=0.02) in a group with scaffold in hypoxia condition, with the highest level on day 4, that is 28,250 ng/ml. The use of enthesis scaffold seeded in adipose-derived mesenchymal stem cells in hypoxic conditions shows a positive trend as regenerative effort of injured enthesis tissue through Scleraxis and Sox9 secretomes induction.