three-dimensional-cell-culture-adipose-derived-stem-cells-hydrogel

Three&#45;Dimensional Cell Culture of Adipose-Derived Stem Cells in a Hydrogel with Photobiomodulation Augmentation

Adipose-derived stem cells (ADSCs), possessing multipotent mesenchymal characteristics akin to stem cells, are frequently employed in regenerative ...

university of Johannesburg

Lung cancer is notably a significant threat when considering worldwide cancer-related deaths. Despite significant advances in treatment modalities, death rates as a result of cancer relapse remain high. Relapse can occur as a result of metastasis. Cancer stem cells (CSCs) have been implicated as an important contributory factor in the development of metastasis. CSCs have the same characteristics as normal stem cells; that is, they can proliferate indefinitely and are capable of both self-renewal and differentiating into specialized cells. The molecular and cellular characteristics of stem cells and CSCs are coded for by cell-specific genes, which can be analyzed by using molecular assays setting the standard to work from. Low-intensity laser irradiation (LILI) has been applied in the treatment of numerous diseases and pathological conditions. LILI has been shown to stimulate proliferation of cells, capillary growth, and cellular metabolism as observed by adenosine triphosphate activation. It has been shown, by using different dosing levels of LILI, to either stimulate or inhibit cellular functions. One treatment strategy used on cancer cells is photodynamic therapy (PDT), in which cancer cells are treated with a photosensitizer (PS) in combination with laser irradiation. PSs are non-toxic by themselves but, with light activation, cause reactive oxygen species generation, which causes cancer cell death. Cell-specific PSs are being developed for future cancer treatment. In this review, we look at the potential effects of LILI and PDT on lung CSCs.

Lung cancer stem cells and low-intensity laser irradiation: a potential future therapy?

The purpose of this in vitro study was to evaluate the effects of low-intensity laser irradiation (LILI) on isolated lung cancer stem cells (CSCs) after several time intervals, using a wavelength of 636 nm and fluences between 5 and 20 J/cm.

Low-Intensity Laser Irradiation at 636 nm Induces Increased Viability and Proliferation in Isolated Lung Cancer Stem Cells.

Photodynamic therapy (PDT) is a promising approach for the treatment of different types of cancer and has been brought into focus for its synergy, compatibility, repeatability, relatively inexpensive cost and it's typically more efficacious nature. Photosensitizers (PSs) play a major role in PDT and are the core of this specific therapy. Al (III) Phthalocyanine Chloride Tetra sulfonic Acid (AlPcSCl) PS is an aromatic macrocyclic metal-based compound that is synthetically derived. It aids in deep tissue penetration due to its far red light activation wavelength, low photo bleaching, increased quantum yields and stability. Lung cancer is a leading cause of cancer related deaths worldwide accounting for approximately 1 in 5 of all cancer-related deaths. In this study, we explored the photochemical properties of AlPcSCl, its uptake into lung cancer, the intracellular localization and photodynamic action on lung cancer (A549 cells). Results indicated that AlPcSCl is a stable PS that localizes in intracellular organelles including the mitochondrion and lysosomes. PDT using AlPcSCl indicated an increase in cell death and decrease in cell proliferation and viability. AlPcSCl showed to be effective in treating lung cancer in vitro, however the resulting PDT efficacy will finally depend on the biological features such as tumour vasculature and tumour specific accumulation when used as a clinical application. It is noted that PDT can be considered as an adjunct therapy until standard protocols for various tumour types along with a relevant PS has been validated.

Effect of dose responses of hydrophilic aluminium (III) phthalocyanine chloride tetrasulphonate based photosensitizer on lung cancer cells.

Cancer stem cells (CSCs) are a leading contributor to lung cancer mortality rates. CSCs are responsible for tumor growth and recurrence through inhibition of drug-induced cell death, decreasing the effect of traditional cancer therapy and photodynamic therapy (PDT). PDT can be improved to successfully treat lung cancer by using gold nanoparticles (AuNPs), due to their size and shape, which have been shown to facilitate drug delivery and retention, along with the targeted antibody (Ab) mediated selection of CSCs. In this study, a nanobioconjugate (NBC) was constructed, using a photosensitizer (PS) (AlPcSCl), AuNPs and Abs. The NBC was characterized, using spectroscopy techniques. Photodynamic effects of the NBC on lung CSCs was evaluated, using biochemical assays 24 h post-irradiation, in order to establish its anticancer effect. Results showed successful conjugation of the nanocomposite. Localization of the NBC was seen to be in integral organelles involved in cell homeostasis. Biochemical responses of lung CSCs treated using AlPcSCl -AuNP and AlPcSCl-AuNP-Ab showed significant cell toxicity and cell death, compared to free AlPcSCl. The PDT effects were enhanced when using the NBC, showing significant lung CSC destruction to the point of eradication.

Effective Gold Nanoparticle-Antibody-Mediated Drug Delivery for Photodynamic Therapy of Lung Cancer Stem Cells.

Given the minimal capacity and sometimes the failure of the mammalian nervous system to regenerate and repair itself after damage, strategies are required to help enhance this regenerative process. Adipose-derived Mesenchymal Stem Cells (ADMSCs) are likely candidates to assist in the recovery process due to their ability to differentiate into neural cells. Successful implementation of this intervention in a clinical setting would increase the rate of recovery following traumatic brain injury.

Potential of Photobiomodulation to Induce Differentiation of Adipose- Derived Mesenchymal Stem Cells into Neural Cells.

Proliferation of osteoblasts is essential for maturation and mineralization of bone matrix. Ossification, the natural phase of bone-forming and hardening is a carefully regulated phase where deregulation of this process may result in insufficient or excessive bone mineralization or ectopic calcification. Osteoblasts can also be differentiated into osteocytes, populating short interconnecting passages within the bone matrix. Over the past few decades, we have seen a significant improvement in awareness and techniques using photobiomodulation (PBM) to stimulate cell function. One of the applications of PBM is the promotion of osteoblast proliferation and maturation. PBM research results on osteoblasts showed increased mitochondrial ATP production, increased osteoblast activity and proliferation, increased and pro-osteoblast expression in the presence of red and NIR radiation. Osteocyte differentiation was also accomplished using blue and green light, showing that different light parameters have various signalling effects. The current review addresses osteoblast function and control, a new understanding of PBM on osteoblasts and its therapeutic impact using various parameters to optimize osteoblast function that may be clinically important. Graphical Abstract.

The Signalling Effects of Photobiomodulation on Osteoblast Proliferation, Maturation and Differentiation: A Review.

Osteoporosis is regarded as the most common chronic metabolic bone condition in humans. In osteoporosis, bone mesenchymal stem cells (MSCs) have reduced cellular function. Regenerative medicine using adipose-derived stem cell (ADSC) transplantation can promote the growth and strength of new bones, improve bone stability, and reduce the risk of fractures. Various methods have been attempted to differentiate ADSCs to functioning specialized cells for prospective clinical application. However, commonly used therapies have resulted in damage to the donor site and morbidity, immune reactions, carcinogenic generation, and postoperative difficulties. Photobiomodulation (PBM) improves ADSC differentiation and proliferation along with reducing clinical difficulties such as treatment failures to common drug therapies and late initiation of treatment. PBM is a noninvasive, nonthermal treatment that encourages cells to produce more energy and to undergo self-repair by using visible green and red and invisible near-infrared (NIR) radiation. The use of PBM for ADSC proliferation and differentiation has been widely studied with multiple outcomes observed due to laser fluence and wavelength dependence. In this article, the potential for differentiating ADSCs into osteoblasts and the various methods used, including biological induction, chemical induction, and PBM, will be addressed. Likewise, the optimal laser parameters that could improve the proliferation and differentiation of ADSC, translating into clinical success, will be commented on.

Photobiomodulation: An Effective Approach to Enhance Proliferation and Differentiation of Adipose-Derived Stem Cells into Osteoblasts.

Regenerative medicine uses undifferentiated adipose-derived mesenchymal stem cells (ADMSCs) to differentiate into multiple cell types. Photobiomodulation (PBM) is a rapidly growing treatment for pain and inflammation reduction, as well as tissue healing. PBM's efficacy is dependent on wavelength and energy dosage. Red (600-700 nm) and near-infrared (780-1100 nm) wavelengths have been shown to promote cell proliferation. Light wavelengths such as green (495 nm-570 nm) have been found to influence ADMSC differentiation. The initiation of ADMSC proliferation and differentiation requires physiologically relevant levels of reactive oxygen species (ROS), while increased levels inhibit self-renewal. Stem cell differentiation is guided by mitochondrial metabolism, where an increased mitochondrial membrane potential (MMP) is associated with higher in vitro differentiation capacity. ADMSCs must home to and accumulate at the sites of injury in regenerative medicine, so cell homing is critical. The aim of this in vitro study was to compare consecutive NIR (825 nm) and green (525 nm) applications on ADMSC morphology and physiology with the possibility that multiple wavelengths could lead to a combination of the two effects. The results showed that concurrent use of NIR-green irradiation significantly stimulated ADMSC proliferation, increasing population density and cellular ATP. Furthermore, NIR-green showed a time dependent increase in ROS production and was significantly higher at 7 days. Consecutive NIR-green irradiation significantly increased MMP and was most effective at facilitating ADMSC migration over time. Findings suggest that with consecutive NIR and green irradiation, the ADMSCs can rapidly proliferate, but can also be modulated for regenerative purposes.

Single and consecutive application of near-infrared and green irradiation modulates adipose derived stem cell proliferation and affect differentiation factors.

Cancer stem cells (CSCs) are considered to contribute to the recurrence of lung cancer due to their stem-like nature and the involvement of genetic markers associated with drug efflux, regeneration and metastases. Photodynamic therapy (PDT) is a cost-effective and non-invasive therapeutic application that can act as an alternative therapy for lung cancer when considering CSC involvement. Stem-like cells derived from the A549 lung cancer cell line, positive for CD133, CD56 and CD44 antigen markers, were characterized, intracellular localization of aluminium (III) phthalocyanine chloride tetrasulphonate (AlPcSCl) determined and its anti-cancer PDT effects were evaluated. Results confirmed that isolated cells were stem cell-like and subcellular localization of AlPcSCl in integral organelles involved in cell homeostasis supported the destruction of CSC. AlPcSCl's effectivity was demonstrated with CSC eradication showing a significant increase in cytotoxicity and cell death via apoptosis, caused by a decrease in mitochondrial membrane potential. PDT could serve as a palliative treatment for lung cancer and improve prognosis by elimination of lung CSCs.

Aluminium (III) phthalocyanine chloride tetrasulphonate is an effective photosensitizer for the eradication of lung cancer stem cells.

Neuronal cells can be generated from adipose-derived stem cells (ADSCs) through biological or chemical inducers. Research has shown that this process may be optimized by the introduction of laser irradiation in the form of photobiomodulation (PBM) to cells. This  study is aimed at generating neuronal-like cells with inducers, chemical or biological, and at furthermore treating these transdifferentiating cells with consecutive PBM of a 525 nm green (G) laser and 825 nm near-infrared (NIR) laser light with a fluence of 10 J/cm. Cells were exposed to induction type 1 (IT1): 3-isobutyl-1-methylxanthine (IBMX) (0.5 mM)+indomethacin (200 M)+insulin (5 g/ml) for 14 days, preinduced with -mercaptoethanol (BME) (1 mM) for two days, and then incubated with IT2: -hydroxyanisole (BHA) (100 M)+retinoic acid (RA) (10-6 M)+epidermal growth factor (EGF) (10 ng/ml)+basic fibroblast growth factor (bFGF) (10 ng/ml) for 14 days and preinduced with -mercaptoethanol (BME) (1 mM) for two days and then incubated with indomethacin (200 M)+RA (1 M)+forskolin (10 M) for 14 days. The results were evaluated through morphological observations, viability, proliferation, and migration studies, 24 h, 48 h, and 7 days post-PBM. The protein detection of an early neuronal marker, neuron-specific enolase (NSE), and late, ciliary neurotrophic factor (CNTF), was determined with enzyme-linked immunosorbent assays (ELISAs). The genetic expression was also explored through real-time PCR. Results indicated differentiation in all experimental groups; however, cells that were preinduced showed higher proliferation and a higher differentiation rate than the group that was not preinduced. Within the preinduced groups, results indicated that cells treated with IT2 and consecutive PBM upregulated differentiation the most morphologically and physiologically.

Promoting Immortalized Adipose-Derived Stem Cell Transdifferentiation and Proliferation into Neuronal-Like Cells through Consecutive 525 nm and 825 nm Photobiomodulation.

Lung cancer remains the leading threat of death globally, killing more people than colon, breast, and prostate cancers combined. Novel lung cancer treatments are being researched because of the ineffectiveness of conventional cancer treatments and the failure of remission. Photodynamic therapy (PDT), a cancer treatment method that is still underutilized, is a sophisticated cancer treatment that shows selective destruction of malignant cells via reactive oxygen species production. PDT has been extensively studied  and clinically. Various PDT strategies have been shown to be effective in the treatment of lung cancer. PDT has been shown in clinical trials to considerably enhance the quality of life and survival in individuals with incurable malignancies. Furthermore, PDT, in conjunction with the use of nanoparticles, is currently being researched for use as an effective cancer treatment, with promising results. PDT and the new avenue of nanoPDT, which are novel treatment options for lung cancer with such promising results, should be tested in clinical trials to determine their efficacy and side effects. In this review, we examine the status and future potentials of nanoPDT in lung cancer treatment.

Photodynamic therapy of lung cancer, where are we?

Nanotechnology-based phototherapies have drawn interest in the fight against cancer because of its noninvasiveness, high flexibility, and precision in terms of cancer targeting and drug delivery based on its surface properties and size. Phototherapy has made remarkable development in recent decades. Approaches to phototherapy, which utilize nanomaterials or nanotechnology have emerged to contribute to advances around nanotechnologies in medicine, particularly for cancers. A brief overviews of the development of photodynamic therapy as well as its mechanism in cancer treatment is provided. We emphasize the design of novel nanoparticles utilized in photodynamic therapy while summarizing the representative progress during the recent years. Finally, to forecast important future research in this area, we examine the viability and promise of photodynamic therapy systems based on nanoparticles in clinical anticancer treatment applications and briefly make mention of the elimination of all reactive metabolites pertaining to nano formulations inside living organisms providing insight into clinical mechanistic processes. Future developments and therapeutic prospects for photodynamic treatments are anticipated. Our viewpoints might encourage scientists to create more potent phototherapy-based cancer therapeutic modalities. This article is categorized under: Therapeutic Approaches and Drug Discovery > Nanomedicine for Oncologic Disease.

Nano-phototherapy: Favorable prospects for cancer treatment.

Worldwide, osteoporosis is the utmost predominant degenerative bone condition. Stem cell regenerative therapy using adipose-derived mesenchymal stem cells (ADMSCs) is a promising therapeutic route for osteoporosis. Photobiomodulation (PBM) has sparked considerable international appeal due to its' ability to augment stem cell proliferation and differentiation properties. Furthermore, the differentiation of ADMSCs into osteoblast cells and cellular proliferation effects have been established using a combination of osteogenic differentiation inducers and PBM. This  study applied dexamethasone, β-glycerophosphate disodium, and ascorbic acid as differentiation inducers for osteogenic induction differentiation media. In addition, PBM at a near-infrared (NIR) wavelength of 825 nm, a green (G) wavelength of 525 nm, and the novel combination of both these wavelengths using a single fluence of 5 J/cm had been applied to stimulate proliferation and differentiation effectivity of immortalised ADMSCs into early osteoblasts. Flow cytometry and ELISA were used to identify osteoblast antigens using early and late osteoblast protein markers. Alizarin red Stain was employed to identify calcium-rich deposits by cells within culture. The morphology of the cells was examined, and biochemical assays such as an EdU proliferation assay, MTT proliferation and viability assay, Mitochondrial Membrane Potential assay, and Reactive Oxygen Species assay were performed. The Central Scratch Test determined the cells' motility potential. The investigative outcomes revealed that a combination of PBM treatment and osteogenic differentiation inducers stimulated promising early osteogenic differentiation of immortalised ADMSCs. The NIR-Green PBM combination did appear to offer great potential for immortalised ADMSC differentiation into early osteoblasts amongst selected assays, however, further investigations will be required to establish the effectivity of this novel wavelength combination. This research contributes to the body of knowledge and assists in the establishment of a standard for osteogenic differentiation  utilising PBM.

Enhancing osteogenic differentiation in adipose-derived mesenchymal stem cells with Near Infra-Red and Green Photobiomodulation.

Regenerative medicine has developed as a promising discipline that utilizes stem cells to address limitations in traditional therapies, using innovative techniques to restore and repair damaged organs and tissues. One such technique is the generation of three-dimensional (3D) organoids in stem cell therapy. Organoids are 3D constructs that resemble specific organs' structural and functional characteristics and are generated from stem cells or tissue-specific progenitor cells. The use of 3D organoids is advantageous in comparison to traditional two-dimensional (2D) cell culture by bridging the gap between in vivo and in vitro research. This review aims to provide an overview of the advancements made towards regenerative medicine using stem cells to generate organoids, explore the techniques used in generating 3D organoids and their applications and finally elucidate the challenges and future directions in regenerative medicine using 3D organoids.

Anine Crous

Laser Research Centre,
Faculty of Health Science,
Laser Research Centre, Faculty of Health Science

university of Johannesburg

Three-Dimensional Cell Culture of Adipose-Derived Stem Cells in a Hydrogel with Photobiomodulation Augmentation

Anine Crous

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university of Johannesburg

Three-Dimensional Cell Culture of Adipose-Derived Stem Cells in a Hydrogel with Photobiomodulation Augmentation

Laser Research Centre,
Faculty of Health Science,
Laser Research Centre, Faculty of Health Science