Outcomes In Clinical Applications Research Articles

Human Brain Organoid: A Versatile Tool for Modeling Neurodegeneration Diseases and for Drug Screening.

Clinical trials serve as the fundamental prerequisite for clinical therapy of human disease, which is primarily based on biomedical studies in animal models. Undoubtedly, animal models have made a significant contribution to gaining insight into the developmental and pathophysiological understanding of human diseases. However, none of the existing animal models could efficiently simulate the development of human organs and systems due to a lack of spatial information; the discrepancy in genetic, anatomic, and physiological basis between animals and humans limits detailed investigation. Therefore, the translational efficiency of the research outcomes in clinical applications was significantly weakened, especially for some complex, chronic, and intractable diseases. For example, the clinical trials for human fragile X syndrome (FXS) solely based on animal models have failed such as mGluR5 antagonists. To mimic the development of human organs more faithfully and efficiently translate in vitro biomedical studies to clinical trials, extensive attention to organoids derived from stem cells contributes to a deeper understanding of this research. The organoids are a miniaturized version of an organ generated in vitro, partially recapitulating key features of human organ development. As such, the organoids open a novel avenue for in vitro models of human disease, advantageous over the existing animal models. The invention of organoids has brought an innovative breakthrough in regeneration medicine. The organoid-derived human tissues or organs could potentially function as invaluable platforms for biomedical studies, pathological investigation of human diseases, and drug screening. Importantly, the study of regeneration medicine and the development of therapeutic strategies for human diseases could be conducted in a dish, facilitating in vitro analysis and experimentation. Thus far, the pilot breakthrough has been made in the generation of numerous types of organoids representing different human organs. Most of these human organoids have been employed for in vitro biomedical study and drug screening. However, the efficiency and quality of the organoids in recapitulating the development of human organs have been hindered by engineering and conceptual challenges. The efficiency and quality of the organoids are essential for downstream applications. In this article, we highlight the application in the modeling of human neurodegenerative diseases (NDDs) such as FXS, Alzheimer's disease (AD), Parkinson's disease (PD), and autistic spectrum disorders (ASD), and organoid-based drug screening. Additionally, challenges and weaknesses especially for limits of the brain organoid models in modeling late onset NDDs such as AD and PD., and future perspectives regarding human brain organoids are addressed.

EFFECT OF Β-TRICALCIUM PHOSPHATE ELEMENTAL IMPURITIES ON THE ANTIBACTERIAL ACTIVITY OF CHITOSAN–GELATIN–ΒTCP COMPOSITE SCAFFOLD

This study aims to fabricate composite a scaffold using a mixture of chitosan-gelatin and limestone-extracted βTCP for better antibacterial properties. Limestone is abundantly available in nature and can provide calcium in the production of calcium phosphate (CaP) materials. However, the elemental impurities present in limestone-extracted βTCP may affect the properties of the CaP-fabricated scaffolds. βTCP powder could be used to fabricate bone scaffold using calcination and wet precipitation method. For example, the preparation of βTCP powder involved the transformation of calcium carbonate in limestone to calcium hydroxide, followed by a reaction with phosphoric acid and sintering treatment to form βTCP. In order to fabricate the scaffold, chitosan was first dissolved in acetic acid and then mixed with a gelatin-in-water solution, followed by adding βTCP powder (limestone-extracted βTCP and commercial βTCP). Then, the mixture was placed in a mold and freeze-dried. Our method improved the antibacterial activity of chitosan–gelatin–βTCP compared to the chitosan–gelatin scaffold. Limestone-extracted βTCP also exhibited higher inhibitory action against bacteria than commercial βTCP. This method's effectiveness evaluation was confirmed by the measurement of X-ray fluorescence analysis, which showed that βTCP scaffolds had more antibacterial elements than the chitosan–gelatin scaffold. These new research results contribute to enhanced antibacterial properties of the material due to elemental impurities in βTCP, especially the limestone-extracted βTCP. However, further evaluation on controlling βTCP impurities is essential for better outcomes in clinical applications.

Albumin Binder-Conjugated Fibroblast Activation Protein Inhibitor Radiopharmaceuticals for Cancer Therapy.

Fibroblast activation protein (FAP) has become an attractive target for diagnosis and therapy, and a series of FAP inhibitor (FAPI)-based radiotracers has been developed and had excellent performance for diagnosis outcomes in clinical applications. Yet, their fast clearance and insufficient tumor retention have hampered their further clinical application in cancer treatment. In this study, we developed 2 albumin binder-conjugated FAPI radiotracers, TEFAPI-06 and TEFAPI-07. They were derived from FAPI-04 and were optimized by conjugating 2 types of well-studied albumin binders, 4-(p-iodophenyl) butyric acid moiety (TEFAPI-06) and truncated Evans blue moiety (TEFAPI-07), to try to overcome the above limitations at the expense of prolonging the blood circulation. Methods: TEFAPI-06 and TEFAPI-07 were synthesized and labeled with 68Ga, 86Y, and 177Lu successfully. A series of cell assays was performed to identify the binding affinity and FAP specificity invitro. PET imaging, SPECT imaging, and biodistribution studies were performed to evaluate the pharmacokinetics in pancreatic cancer patient-derived xenograft (PDX) animal models. The cancer treatment efficacy of 177Lu-TEFAPI-06 and 177Lu-TEFAPI-07 were evaluated in pancreatic cancer PDX-bearing mice. Results: The binding affinities (dissociation constants) to FAP of 68Ga-TEFAPI-06 and 68Ga-TEFAPI-07 were 10.16 ± 2.56 nM and 7.81 ± 2.28 nM, respectively, which were comparable with that of 68Ga-FAPI-04. Comparative PET imaging of HT-1080-FAP and HT-1080 tumor-bearing mice and a blocking study showed the FAP-targeting ability invivo of these 2 tracers. Compared with 177Lu-FAPI-04, PET imaging, SPECT imaging, and biodistribution studies of TEFAPI-06 and TEFAPI-07 demonstrated their remarkably enhanced tumor accumulation and retention, respectively. Notable tumor growth inhibition by 177Lu-TEFAPI-06 and 177Lu-TEFAPI-07 were observed, whereas the control group and the group treated by 177Lu-FAPI-04 showed a slight therapeutic effect. Conclusion: Two albumin binder-conjugated FAPI radiopharmaceuticals have been developed and evaluated invitro and invivo. Significantly improved tumor uptake and retention were observed, compared with the original FAPI tracer. Both 177Lu-TEFAPI-06 and 177Lu-TEFAPI-07 showed remarkable growth inhibition of PDX tumors, whereas the side effects were almost negligible, demonstrating that these radiopharmaceuticals are promising for further clinical translational studies.

Design of a sustainable prepolarizing magnetic resonance imaging system for infant hydrocephalus.

The need for affordable and appropriate medical technologies for developing countries continues to rise as challenges such as inadequate energy supply, limited technical expertise, and poor infrastructure persist. Low-field magnetic resonance imaging (LF MRI) is a technology that can be tailored to meet specific imaging needs within such countries. Its low power requirements and the possibility of operating in minimally shielded or unshielded environments make it especially attractive. Although the technology has been widely demonstrated over several decades, it is yet to be shown that it can be diagnostic and improve patient outcomes in clinical applications. We here demonstrate the robustness of prepolarizing MRI (PMRI) technology for assembly and deployment in developing countries for the specific application to infant hydrocephalus. Hydrocephalus treatment planning and management requires only modest spatial resolution, such that the brain can be distinguished from fluid-tissue contrast detail within the brain parenchyma is not essential. We constructed an internally shielded PMRI system based on the Lee-Whiting coil system with a 22-cm diameter of spherical volume. In an unshielded room, projection phantom images were acquired at 113kHz with in-plane resolution of 3mm × 3mm, by introducing gradient fields of sufficient magnitude to dominate the 5000 ppm inhomogeneity of the readout field. The low cost, straightforward assembly, deployment potential, and maintenance requirements demonstrate the suitability of our PMRI system for developing countries. Further improvement in image spatial resolution and contrast of LF MRI will broaden its potential clinical utility beyond hydrocephalus.

Different characteristics of mesenchymal stem cells isolated from different layers of full term placenta.

BackgroundThe placenta is a very attractive source of mesenchymal stem cells (MSCs) for regenerative medicine due to readily availability, non-invasive acquisition, and avoidance of ethical issues. Isolating MSCs from parts of placenta tissue has obtained growing interest because they are assumed to exhibit different proliferation and differentiation potentials due to complex structures and functions of the placenta. The objective of this study was to isolate MSCs from different parts of the placenta and compare their characteristics.MethodsPlacenta was divided into amniotic epithelium (AE), amniotic membrane (AM), chorionic membrane (CM), chorionic villi (CV), chorionic trophoblast without villi (CT-V), decidua (DC), and whole placenta (Pla). Cells isolated from each layer were subjected to analyses for their morphology, proliferation ability, surface markers, and multi-lineage differentiation potential. MSCs were isolated from all placental layers and their characteristics were compared.FindingsSurface antigen phenotype, morphology, and differentiation characteristics of cells from all layers indicated that they exhibited properties of MSCs. MSCs from different placental layers had different proliferation rates and differentiation potentials. MSCs from CM, CT-V, CV, and DC had better population doubling time and multi-lineage differentiation potentials compared to those from other layers.ConclusionsOur results indicate that MSCs with different characteristics can be isolated from all layers of term placenta. These finding suggest that it is necessary to appropriately select MSCs from different placental layers for successful and consistent outcomes in clinical applications.

Scientific assessment on the outcome of clinical application with peripheral nerve graft materials

Objective To discuss how to assess scientifically on the outcome of clinical application with peripheral nerve graft materials. Methods All Pubmed database from 1990 to 2010 were retrieved,and searched the English literatures about the application with peripheral nerve graft materials. The literatures consisted of original clinical research and review excluding animal experiments, repetitive research and irrelevant literatures. The clinical trials data of U.S. was also our target. The information about the safety and effectiveness of peripheral nerve graft materials and related statistical problems were discussed. Results Totally 1578 literatures were identified. Following reading titles and abstracts, we excluded some irrelevant articles. Finally 31 literatures and 2 issue of clinical research from clinical trial data of U.S. were included. After analysis on the literatures, we gained the following results: a remarkable degree of homogeneity among patients can be formed by setting the inclusion and exclusion criteria. For the assessment of proper digital nerve repair, Macknnon- Dellon evaluation is commonly applied, but for the composite nerve, BMRC evaluation is the main method and electromyography can be used as a secondary choice. The safety of peripheral nerve graft materials cannot be evaluated throughout one's life according to the current level of science and technology. It should be evaluated by long-term clinical observation. Randomized clinical trials with random grouping was a gold standard for clinical trials with a good balance and strong comparability. However, non-randomized controlled trials also have an important value. Conclusion It is impossible to make all affected factors homogeneity in a limited timespace conditions of clinical trial. However, we can try our best to keep factors homogeneity to maximum degree by setting the inclusion and exclusion criteria. The scientific assessment of outcome of peripheral nerve repair can be carried out with reasonable and internationally recognized nerve function evaluation methods, strict follow-up time and statistics programme meeting the clinical requirement. Key words: Peripheral nerve; Graft materials; Nerve function evaluation; Literatures assessment

Combined treatment of human MCF-7 breast carcinoma with antibody, cationic lipid and hyaluronic acid using ex vivo assays

The effective targeting of malignant cell surface antigens is essential in cancer therapy. Resistance to treatment and rapid invasion of cancer cells are the main causes of cancer mortality. Despite intense research efforts, treatments often have demonstrated insufficient outcomes in clinical applications. The aim of the present study was to determine whether combined administration of monoclonal antibody (Herceptin ®, trastuzumab) and anti-HER-2 (clone CB11) with hyaluronic acid (HA) and lipoplex (containing lipofectamine (LipA) and plasmid DNA) can produce a synergistic reaction to increase the therapeutic effect of monoclonal antibodies. To assess the treatment response, we cultured a 3-D MCF-7 cell line overexpressing HER-2 and CD44 receptors. The high density 3-D cell aggregation in the hollow fiber bioreactor (HFB) used for the cell culture was monitored with the use of proton magnetic resonance imaging ( 1H MRI). In addition, matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS) was used in combination with HPLC (high performance liquid chromatography) to evaluate structural changes in the proteins contained in treated cells. The study showed that incorporation of antibodies into targeted lipoplex results in more efficient delivery of the complex to tumor cells. The viability of cells decreased mostly due to cellular uptake of lipoplex and binding of the antibodies to the cellular surface receptor. The data also demonstrate that HA could be used to enhance treatment efficacy of trastuzumab and anti-HER-2 (clone CB11) in breast cancer cell cultures.

Photo-toxic effects of 809-nm diode laser and indocyanine green on MDA-MB231 breast cancer cells

Photodynamic therapy (PDT), due to its positive outcomes in clinical applications, easiness of practice and few side effects, is a good candidate for an efficient treatment of cancer. Indocyanine green (ICG), a water-soluble, anionic tricarbocyanine and non-toxic molecule is a promising photosensitive agent for PDT applications on tumor cells. ICG exhibits strong maximum absorption at around 805 nm which will be an advantage for its use in PDT; light at that wavelength can be used to treat deeper tumors. In this study the inhibitory growth effects of ICG-PDT on MDA-MB231 human breast cancer cells were investigated in a time course experiment. Cells were irradiated with a continuous wave diode laser (lambda=809 nm, 60 mW, 24 J cm(-2)). Cell viability was measured by MTT assay 0, 3, 6, 9, 12, 24 and 48h after light irradiation. The results showed that ICG-PDT application exerted its photo-oxidative effect on MDA-MB231 breast cancer cells immediately. Relative cell viability was determined throughout the 48h time course, and a consistent decrease was observed after ICG-PDT applications. In conclusion, ICG when used in combination with near-infrared light showed a very fast (within 3h) and persistent (up to 48h) photo-toxic effect on MDA-MB231 human breast cancer cells.