Disc Cells Research Articles

Harmonization and standardization of nucleus pulposus cell extraction and culture methods.

In vitro studies using nucleus pulposus (NP) cells are commonly used to investigate disc cell biology and pathogenesis, or to aid in the development of new therapies. However, lab-to-lab variability jeopardizes the much-needed progress in the field. Here, an international group of spine scientists collaborated to standardize extraction and expansion techniques for NP cells to reduce variability, improve comparability between labs and improve utilization of funding and resources. The most commonly applied methods for NP cell extraction, expansion, and re-differentiation were identified using a questionnaire to research groups worldwide. NP cell extraction methods from rat, rabbit, pig, dog, cow, and human NP tissue were experimentally assessed. Expansion and re-differentiation media and techniques were also investigated. Recommended protocols are provided for extraction, expansion, and re-differentiation of NP cells from common species utilized for NP cell culture. This international, multilab and multispecies study identified cell extraction methods for greater cell yield and fewer gene expression changes by applying species-specific pronase usage, 60-100 U/ml collagenase for shorter durations. Recommendations for NP cell expansion, passage number, and many factors driving successful cell culture in different species are also addressed to support harmonization, rigor, and cross-lab comparisons on NP cells worldwide.

Characterization of temporomandibular joint articular disc progenitor cell clones.

A critical component of the temporomandibular joint (TMJ) is the fibrocartilage articular disc (AD). Researchers have attempted to regenerate the AD to alleviate TMJ osteoarthritis but alternative cell sources for use in AD regenerative approaches are needed due to insufficient extracellular matrix (ECM) production by total articular disc cells (TACs). Tissue-specific progenitor cells have been identified in many tissues. The aim of the present study was to identify adult multipotent progenitor cells within the AD suitable for regenerative medicine applications. A novel AD progenitor cell population was identified in rhesus macaques. Clonally derived articular disc progenitor cells (ADPs) were isolated using fibronectin differential cell adhesion. ADPs represent between 1 and 3 % of the TAC population and are capable of in vitro expansion beyond 60 population doublings. ADPs were characterized using osteogenic, adipogenic, and fibrochondrogenesis differentiation assays. Clones exhibited phenotypic plasticity, differentiating into osteocytes, adipocytes, and fibrochondrocytes. ECM secretion profiles following fibrochondrogenic differentiation were assessed using immunohistochemistry (IHC), fluorescently activated cell sorting (FACS), total collagen, and glycosaminoglycan (GAG) assays and compared with TACs, articular cartilage progenitor cells (ACPs), tendon progenitor cells (TPCs) and bone-marrow-derived mesenchymal stem cells (BMMSCs). ADP pellet cultures produced a biochemical phenotype similar to native AD tissue, with production of versican (VCAN) and collagen types I, II, III, and VI (COL1, COL2, COL3, COL6). However, clonally derived ADP cell lines produced different amounts of ECM and exhibited different expansion potentials. These findings indicated flexibility in clone selection for potential regenerative strategies to recapitulate native anisotropy.

Gene Therapy for Intervertebral Disc Degeneration: An Overview of Current Strategies and Applications

Intervertebral disc degeneration is a pervasive condition contributing to chronic back pain, affecting up to a third of the population, with risk further increasing with age. It is a significant driver of disability for millions of Americans and others worldwide. The standard of care today is reliant on symptomatic treatment rather than addressing the root cause of disease. Surgical interventions alter the structural integrity and biomechanics of the spine, often leading to loss of function and motion, and post-operative complications. This is the basis for innovation in novel biologic treatments, including gene therapy, which aims to reestablish the optimal balance between matrix catabolism and anabolism within pathologically degenerating disc cells. This review will cover the significant advances that have led to identification of target therapeutic genes combined with regulated expression of the therapeutic transgene and successful systems for gene delivery into cells. Recent advances in viral and non-viral vectors for gene transfer, silencing of genes by RNA interference, editing of genes by clustered regularly interspaced short palindromic repeats, and modifying mammalian target of rapamycin signaling pathwayS offer promising treatment avenues. Clinical translation of these approaches, however, will require further investigation of the pathological basis of disc degeneration in addition to systematic safety measures for the adoption of gene therapy.

سطارة جديدة لزراعة البسلة على مصاطب

Until now, the traditional method for planting pea seeds in Egypt is manual because of the scarcity of planting machines. Therefore, this study aims to provide and evaluate a new seed drill for planting pea seeds on a raised bed in silt-clay loam soil. This seed drill consists of the frame, seeds hopper, seed metering device, transmission system, and covering unit. Laboratory tests on pea seeds were conducted to determine the seeds’ physical and mechanical properties. Field trials were carried out under the following parameters; four forward speeds (‎‎0.79, 0.98, 1.28, and 1.64 m s–1), ‎three disc cell capacities (1, 2, and ‎‎3 seeds per cell), two different cells shapes (circular, and a rectangle with semicircle end), and three distances between rows (‎7.5, 10, and 15 cm) to assess the performance of the seed drill on fuel consumption, specific energy, slip ratio, seeds damaged‎, germination ratio, plants number per hill, longitudinal scattering, and pea pods yield. The results revealed that the optimum performance of the seed drill was achieved at a forward speed ranging from 0.79 to 0.98 m s–1, using a disc cell capacity of ‎‎2 seeds, a circular cell shape, and a distance between rows of 10 cm.

Transgenerational inheritance of wing development defects in Drosophila melanogaster induced by cadmium

The transgenerational inheritance of phenotype induced by environmental factors is a new focus in epigenetic research. In this study, Drosophila melanogaster (F0) was cultured in the medium containing cadmium (Cd, 4.5 mg/kg) from eggs to adults, and offspring (F1-F4) were continuously kept in standard medium (without cadmium). The phenotype analysis showed that cadmium induced developmental defects on wings and apoptosis in the wing disc cells of Drosophila (F0). The wing defects were transmitted for at least four generations even without Cd afterwards. And the effect on the mRNA expression of wing development related genes (shg, omb, F-actin, Mekk1) can be maintained for at least two or three generations. More importantly, under cadmium stress, the post-translational modification (PTM) on the histones H3K4me3 in the third instar larvae and ovaries or testes of adult flies increased significantly, while the levels of H3K9me3 and H3K27me3 decreased significantly. The expression of histone methylation related genes (dSet-1, ash1, Lsd1) increased significantly and these changes can be transmitted to offspring from one or two generations in ovaries or testes. These results suggest that the phenotypic defects of wings caused by cadmium can be inherited to the offspring, and this transgenerational inheritance effect may be related to the epigenetic regulation of histone methylation. Therefore, the adaptability of offspring should be considered when evaluating the toxicity and environmental risk of cadmium.

Optic disc, macula and ganglion cell layer measurements obtained with optical coherence tomography in patients with thyroid disorder

Purpose: The aim of this study was to compare retinal nerve layer thickness, central macula thickness and ganglion cell layer measurements using optical coherence tomography (OCT) between thyroid patients and age- and sex-matched healthy controls.
 Materials and Methods: Eighty eyes of 80 patients in the thyroid patient group and 48 eyes of 48 patients in the control group were included in the study. After evaluating the exclusion criteria, visual acuity, anterior segment biomicroscopy, intraocular pressure, fundoscopy and spectral field optic coherence tomography (SF-OCT) imaging tests were performed on all patients included in the study. These data have been recorded. Central macular thickness (CMT), retinal nerve fiber layer (RNFL) and ganglion cell complex (GCC) measurements were evaluated by statistical methods for both groups.
 Results: There was no statistically significant difference between the demographic data of the groups included in the study. IOP measurements were found to be statistically significantly higher in the patient group compared to the control group. CMT values were 258.2±16.1 µm in the patient group and 273.1±23.8 µm in the control group, and statistically significant differences were observed between the groups. 
 Conclusion: The findings supports that changes in the optic nerve and macula develop before vision loss in TO patients compared to the healthy control group. We can follow TO patients more objectively and reliably and make treatment planning more accurately by adding OCT, which is a noninvasive test, to our clinical practice, since structural changes develop before functional deterioration in TO patients.

The role of oxidative stress in intervertebral disc cellular senescence.

With the aggravation of social aging and the increase in work intensity, the prevalence of spinal degenerative diseases caused by intervertebral disc degeneration(IDD)has increased yearly, which has driven a heavy economic burden on patients and society. It is well known that IDD is associated with cell damage and degradation of the extracellular matrix. In recent years, it has been found that IDD is induced by various mechanisms (e.g., genetic, mechanical, and exposure). Increasing evidence shows that oxidative stress is a vital activation mechanism of IDD. Reactive oxygen species (ROS) and reactive nitrogen species (RNS) could regulate matrix metabolism, proinflammatory phenotype, apoptosis, autophagy, and aging of intervertebral disc cells. However, up to now, our understanding of a series of pathophysiological mechanisms of oxidative stress involved in the occurrence, development, and treatment of IDD is still limited. In this review, we discussed the oxidative stress through its mechanisms in accelerating IDD and some antioxidant treatment measures for IDD.

O-Vanillin Modulates Cell Phenotype and Extracellular Vesicles of Human Mesenchymal Stem Cells and Intervertebral Disc Cells.

Human mesenchymal stem cell (hMSC) and extracellular vesicle (EV) therapy is a promising treatment for discogenic low back pain (LBP). Although promising, major obstacles remain to be overcome. Cellular senescence reduces self-renewal and multipotent potentials, and the senescence-associated secretory phenotype creates an inflammatory environment negatively affecting tissue homeostasis. Reducing senescence could therefore improve regenerative approaches. Ortho-Vanillin (o-Vanillin) has senolytic activity and anti-inflammatory properties and could be a valuable supplement to MSC and EV therapy. Here, we used direct co-culture experiments to evaluate proteoglycan synthesis, inflammatory mediators, and senescent cells in the presence or absence of o-Vanillin. EV release and transfer between hMSCs and intervertebral disc cells (DCs) was examined, and the effect on hMSC differentiation and DC phenotype was evaluated in the presence and absence of o-Vanillin. This study demonstrates that o-Vanillin affects cell communication, enhances hMSC differentiation and improves DC phenotype. Co-cultures of DCs and hMSCs resulted in increased proteoglycan synthesis, a decreased number of senescent cells and decreased release of the cytokines IL6 and 8. Effects that were further enhanced by o-Vanillin. o-Vanillin profoundly increased EV release and/or uptake by hMSCs and DCs. DC markers were significantly upregulated in both cell types in response to conditioned media of o-Vanillin treated donor cells. Collectively, this study demonstrates that o-Vanillin affects hMSC and DC crosstalk and suggests that combining hMSCs and senolytic compounds may improve the outcome of cell supplementation and EV therapy for LBP.

The role of ageing and oxidative stress in intervertebral disc degeneration.

Intervertebral disc degeneration (IDD) is the primary cause of intervertebral disc (IVD) disease. With the increased ageing of society, an increasing number of patients are plagued by intervertebral disc disease. Ageing not only accelerates the decreased vitality and functional loss of intervertebral disc cells but also increases intracellular oxidative stress. Moreover, the speed of intervertebral disc ageing is also linked to high levels of reactive oxygen species (ROS) production. Not only is the production of ROS increased in ageing intervertebral disc cells, but antioxidant levels in degenerative intervertebral discs also decrease. In addition to the intervertebral disc, the structural components of the intervertebral disc matrix are vulnerable to oxidative damage. After chronic mitochondrial dysfunction, ROS can be produced in large quantities, while autophagy can eliminate these impaired mitochondria to reduce the production of ROS. Oxidative stress has a marked impact on the occurrence of IDD. In the future, IDD treatment is aiming to improve oxidative stress by regulating the redox balance in intervertebral disc cells. In summary, ageing and oxidative stress promote the degeneration of IVD, but further basic and clinical trials are needed to determine how to treat oxidative stress. At present, although there are many in-depth studies on the relationship between oxidative stress and degeneration of intervertebral disc cells, the specific mechanism has not been elucidated. In this paper, the main causes of intervertebral disc diseases are studied and summarized, and the impact of oxidative stress on intervertebral disc degeneration is studied.

Targeting Ferroptosis Holds Potential for Intervertebral Disc Degeneration Therapy.

Intervertebral disc degeneration (IVDD) is a common pathological condition responsible for lower back pain, which can significantly increase economic and social burdens. Although considerable efforts have been made to identify potential mechanisms of disc degeneration, the treatment of IVDD is not satisfactory. Ferroptosis, a recently reported form of regulated cell death (RCD), is characterized by iron-dependent lipid peroxidation and has been demonstrated to be responsible for a variety of degenerative diseases. Accumulating evidence suggests that ferroptosis is implicated in IVDD by decreasing viability and increasing extracellular matrix degradation of nucleus pulposus cells, annulus fibrosus cells, or endplate chondrocytes. In this review, we summarize the literature regarding ferroptosis of intervertebral disc cells and discuss its molecular pathways and biomarkers for treating IVDD. Importantly, ferroptosis is verified as a promising therapeutic target for IVDD.

Regulated cell death: Implications for intervertebral disc degeneration and therapy.

Intervertebral disc degeneration (IDD) is the major source of chronic low back pain. As a controllable biological process, regulated cell death (RCD) extensively participates in the pathogenesis of IDD. Nevertheless, the initiation and progression of RCD remain unclear, and more importantly, the interaction between different RCD modes during IDD and therapy is far from well understood. Literature search was performed using "regulated cell death AND intervertebral disc degeneration" in PubMed, Embase, and Web of Science. Meanwhile, relevant findings have been reviewed and quoted. In this review, we discuss the inducing factors of IDD, various modes of RCD in intervertebral disc, the interactions between different RCD modes, as well as the obstacles to achieve disc regeneration. Meanwhile, the research gaps and perspective in studies that targeting RCD are also presented. Increasing evidence demonstrated the presence of different RCD modes in intervertebral disc during the progression of IDD. RCD in the resident disc cells is probably induced by the multiple factors such as abnormal mechanical loading, nutritional imbalance, inflammation microenvironment, circadian rhythm changes, withdraw of hormones, and other biomechanical factors. A better understanding of the fundamental mechanisms and the interactions between different RCD modes might contribute to the rescuing of disc degeneration and development of promising therapeutics. The Translational potential of this article. This review aims to demonstrate a better understanding of the fundamental mechanisms governing RCD, which might contribute to the rescuing of disc degeneration and to the development of promising therapeutics in a clinical setting.

Pharmacological Effects of Resveratrol in Intervertebral Disc Degeneration: A Literature Review.

Intervertebral disc degeneration (IDD) is a high incidence disease of musculoskeletal system that often leads to stenosis, instability, pain and even deformity of the spinal segments. IDD is an important cause of discogenic lower back pain and often leads to large economic burden to families and society. Currently, the treatment of IDD is aimed at alleviating symptoms rather than blocking or reversing pathological progression of the damaged intervertebral disc. Resveratrol (RSV) is a polyphenol phytoalexin first extracted from the Veratrum grandiflflorum O. Loes and can be found in various plants and red wine. Owing to the in-depth study of pharmacological mechanisms, the therapeutic potential of RSV in various diseases such as osteoarthritis, neurodegenerative diseases, cardiovascular diseases and diabetes have attracted the attention of many researchers. RSV has anti-apoptotic, anti-senescent, anti-inflammatory, anti-oxidative, and anabolic activities, which can prevent further degeneration of intervertebral disc cells and enhance their regeneration. With high safety and various biological functions, RSV might be a promising candidate for the treatment of IDD. This review summarizes the biological functions of RSV in the treatment of IDD and to facilitate further research.

What's New in Musculoskeletal Basic Science.

What's New in Musculoskeletal Basic Science.

The Involvement of Cell-Type-Specific Glycans in Hydra Temporary Adhesion Revealed by a Lectin Screen

Hydra is a freshwater solitary polyp, capable of temporary adhesion to underwater surfaces. The reversible attachment is based on an adhesive material that is secreted from its basal disc cells and left behind on the substrate as a footprint. Despite Hydra constituting a standard model system in stem cell biology and tissue regeneration, few studies have addressed its bioadhesion. This project aimed to characterize the glycan composition of the Hydra adhesive, using a set of 23 commercially available lectins to label Hydra cells and footprints. The results indicated the presence of N-acetylglucosamine, N-acetylgalactosamine, fucose, and mannose in the adhesive material. The labeling revealed a meshwork-like substructure in the footprints, implying that the adhesive is mainly formed by fibers. Furthermore, lectins might serve as a marker for Hydra cells and structures, e.g., many labeled as glycan-rich nematocytes. Additionally, some unexpected patterns were uncovered, such as structures associated with radial muscle fibers and endodermal gland cells in the hypostome of developing buds.

Innovative immune mechanisms and antioxidative therapies of intervertebral disc degeneration.

Intervertebral disc degeneration (IDD) is the basic pathological process of many degenerative diseases of the spine, characterized by series of symptoms, among which low back pain (LBP) is the most common symptom that patients suffer a lot, which not only makes patients and individual families bear a huge pain and psychological burden, but also consumes a lot of medical resources. IDD is usually thought to be relevant with various factors such as genetic predisposition, trauma and aging, and IDD progression is tightly relevant with structural and functional alterations. IDD processes are caused by series of pathological processes, including oxidative stress, matrix decomposition, inflammatory reaction, apoptosis, abnormal proliferation, cell senescence, autophagy as well as sepsis process, among which the oxidative stress and inflammatory response are considered as key link in IDD. The production and clearance of ROS are tightly connected with oxidative stress, which would further simulate various signaling pathways. The phenotype of disc cells could change from matrix anabolism-to matrix catabolism- and proinflammatory-phenotype during IDD. Recent decades, with the relevant reports about oxidative stress and inflammatory response in IDD increasing gradually, the mechanisms researches have attracted much more attention. Consequently, this study focused on the indispensable roles of the oxidative stress and inflammatory response (especially macrophages and cytokines) to illustrate the origin, development, and deterioration of IDD, aiming to provide novel insights in the molecular mechanisms as well as significant clinical values for IDD.

Emerging role and therapeutic implication of mTOR signalling in intervertebral disc degeneration.

Intervertebral disc degeneration (IDD), an important cause of chronic low back pain (LBP), is considered the pathological basis for various spinal degenerative diseases. A series of factors, including inflammatory response, oxidative stress, autophagy, abnormal mechanical stress, nutritional deficiency, and genetics, lead to reduced extracellular matrix (ECM) synthesis by intervertebral disc (IVD) cells and accelerate IDD progression. Mammalian target of rapamycin (mTOR) is an evolutionarily conserved serine/threonine kinase that plays a vital role in diverse degenerative diseases. Recent studies have shown that mTOR signalling is involved in the regulation of autophagy, oxidative stress, inflammatory responses, ECM homeostasis, cellular senescence, and apoptosis in IVD cells. Accordingly, we reviewed the mechanism of mTOR signalling in the pathogenesis of IDD to provide innovative ideas for future research and IDD treatment.

Encapsulation of Manganese Porphyrin in Chondroitin Sulfate-A Microparticles for Long Term Reactive Oxygen Species Scavenging.

Oxidative stress due to excess reactive oxygen species (ROS) is related to many chronic illnesses including degenerative disc disease and osteoarthritis. MnTnBuOE-2-PyP5+ (BuOE), a manganese porphyrin analog, is a synthetic superoxide dismutase mimetic that scavenges ROS and has established good treatment efficacy at preventing radiation-induced oxidative damage in healthy cells. BuOE has not been studied in degenerative disc disease applications and only few studies have loaded BuOE into drug delivery systems. The goal of this work is to engineer BuOE microparticles (MPs) as an injectable therapeutic for long-term ROS scavenging. Methacrylated chondroitin sulfate-A MPs (vehicle) and BuOE MPs were synthesized via water-in-oil polymerization and the size, surface morphology, encapsulation efficiency and release profile were characterized. To assess long term ROS scavenging of BuOE MPs, superoxide scavenging activity was evaluated over an 84-day time course. In vitro cytocompatibility and cellular uptake were assessed on human intervertebral disc cells. BuOE MPs were successfully encapsulated in MACS-A MPs and exhibited a slow-release profile over 84days. BuOE maintained high potency in superoxide scavenging after encapsulation and after 84days of incubation at 37°C as compared to naked BuOE. Vehicle and BuOE MPs (100µg/mL) were non-cytotoxic on nucleus pulposus cells and MPs up to 23µm were endocytosed. BuOE MPs can be successfully fabricated and maintain potent superoxide scavenging capabilities up to 84-days. In vitro assessment reveals the vehicle and BuOE MPs are not cytotoxic and can be taken up by cells. The online version contains supplementary material available at 10.1007/s12195-022-00744-w.

Mass Purification Protocol for Drosophila melanogaster Wing Imaginal Discs: An Alternative to Dissection to Obtain Large Numbers of Disc Cells.

Simple SummaryDrosophila melanogaster, also known as the fruit fly, is a widely used organism model, especially for genetic studies or as a model for pathologies. The Drosophila genome is well known and conserved within humans, thus allowing biologists to obtain numerous mutants and transgenic flies from the fruit flies. Gene function studies at the cellular and molecular levels are often performed using extracts of larval tissues. Due to their small size, it is difficult to dissect substantial amounts of these tissues for performing genomic or proteomic experiments. This paper develops a simple method to purify larval tissues en masse. This protocol preserves tissue integrity in the same way as manual dissection; the protocol is achievable by individual researchers and allows the purification of different samples simultaneously.Drosophila melanogaster imaginal discs are larval internal structures that become the external organs of the adult. They have been used to study numerous developmental processes for more than fifty years. Dissecting these imaginal discs for collection is challenging, as the size of third-instar larvae organs is typically less than 1 mm. Certain experimental applications of the organs require many cells, which requires researchers to spend several hours dissecting them. This paper proposes an alternative to dissection in the form of a mass enrichment protocol. The protocol enables the recovery of many wing imaginal discs by grinding large quantities of third-instar larvae and separating the organs using filtration and a density gradient. The wing imaginal discs collected with this protocol in less than three hours are as well preserved as those collected by dissection. The dissociation and filtration of the extract allow the isolation of a large amount of wing imaginal disc cells.

Researches on Stem and Progenitor Cells in Intervertebral Discs: An Analysis of the Scientific Landscape.

Low back pain (LBP) is a common clinical symptom, and the prevalence is ranged from 60% to 70%. With the deepening of basic research, the development of intervertebral disc regeneration-oriented cell therapy, especially stem and progenitor cells therapy, showed good research prospects and was expected to become new methods of treatment for LBP. Our study is aimed at analyzing the scientific output of stem and progenitor cells in intervertebral discs and at driving future research into new publications. Researches focused on this file were searched from the Science Citation Index Expanded (SCI-E) of the Web of Science (WOS) core collection database and were screened according to inclusion criteria. We evaluated and visualized the results, including annual publications, citations, authors, organizations, countries, research directions, funds, and journals by bibliometric website, VOSviewer, and Citespace softwares on May 27, 2022. A total of 450 original articles and reviews were included, and the overall trend of the number of publications rapidly increased. In worldwide, China and the USA were the leading countries for research production. The retrieved 450 publications received 14322 citations, with an average of 31.83 citations and an H-index of 62. The most high-yield author, organization, country, research directions, funds, and journals were Chen QX from Zhejiang University, Zhejiang University, China, Cell Biology, National Natural Science Foundation of China, and Spine, respectively. Keywords cluster analysis showed the research hotspots in the future, including “human intervertebral disc”, “adipose-derived mesenchymal stem cell”, “intervertebral disc degeneration”, “degenerative disc model”, “nucleus pulposus regeneration”, “human cartilage”, “3d culture”, “shrinkage-free preparation”, and “polylactide disc”. Furthermore, with accumulating evidence demonstrating the role of stem and progenitor cells in intervertebral discs, “microenvironment”, “activation”, “intervertebral disc degeneration”, and “oxidative stress” are becoming the research frontiers and trends.

940 nm diode laser induced differentiation of human adipose derived stem cells to temporomandibular joint disc cells

BackgroundTemporomandibular disorder (TMD) refers to a group of disorders that affect temporomandibular joint (TMJ) and its associated muscles with very limited treatment options. Stem cell research is emerging as one of the promising fields in the treatment of degenerative diseases. The ability of human adipose derived stem cells to differentiate into many cell types is driving special interest in several disease management strategies. Photobiomodulation has enhanced the role of these stem cells through their ability to promote cell proliferation and differentiation. Hence, this study examined the differentiation potential of human adipose derived stem cells (ADSCs) into fibroblasts and chondrocytes using a 940 nm diode laser for possible TMD therapy.Materials and methodsADSCs were cultured at different seeding densities and for different time intervals. After irradiation at 24, 48, 72 h, 1, 2 and 3 weeks, ADSC viability and morphological changes were assessed in groups with and without basic fibroblast growth factor. Additionally, the level of adenosine triphosphate (ATP) in the cells was also recorded. The differentiated fibroblasts and chondrocytes were characterized with flow cytometry and immunofluorescence techniques, at 1- and 2-weeks post-irradiation.ResultsIncreased ATP proliferation and cell viability above 90% were observed in all post-irradiation experimental groups. Post irradiation results from flow cytometry and immunofluorescence at 1- and 2‐weeks confirmed the expression of chondrogenic and fibroblastic cell surface markers.ConclusionThis study describes stimulatory techniques utilized to differentiate ADSCs into fibroblastic and chondrogenic phenotypes using diode lasers at 940 nm. The study proposes a new treatment model for patients with degenerative disc diseases of the TMJ. The study will offer new possibilities in tissue engineering and TMJ disc management through photobiomodulation of ADSCs using a 940 nm diode laser.