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Novel method boosts MSC efficacy for articular cartilage repair

October 18, 2024
in Health
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Researchers from the Crucial Analytics for Manufacturing Customized-Drugs (CAMP) Interdisciplinary Analysis Group (IRG) at Singapore-MIT Alliance for Analysis and Know-how (SMART), MIT’s analysis enterprise in Singapore, alongside collaborators from the Nationwide College of Singapore Tissue Engineering Programme (NUSTEP), have developed a novel methodology to boost the power of mesenchymal stromal cells (MSCs) to generate cartilage tissue by including ascorbic acid (AA) throughout MSC enlargement. The analysis additionally found that micro magnetic resonance relaxometry (µMRR), a novel course of analytical software (PAT) developed by SMART CAMP, can be utilized as a speedy, label-free process-monitoring software for the standard enlargement of MSCs.

Micro magnetic resonance relaxometry (µMRR) is a speedy, label-free process-monitoring software for the enlargement of MSCs. Picture Credit score: SMART

Articular cartilage, a connective tissue that protects the bone ends in joints, can degenerate as a result of damage, age, or arthritis, resulting in important joint ache and incapacity. Particularly in international locations – resembling Singapore – which have an energetic, ageing inhabitants, articular cartilage degeneration is a rising ailment that impacts an growing variety of individuals. Autologous chondrocyte implantation is at present the one FDA-approved cell-based remedy for articular cartilage accidents, however it’s pricey, time-intensive, and requires a number of remedies. MSCs are a sexy and promising different as they’ve proven good security profiles for transplantation. Nevertheless, medical use of MSCs is restricted as a result of inconsistent remedy outcomes arising from components resembling donor-to-donor variability, variation amongst cells throughout cell enlargement and unstandardized MSC manufacturing protocols.

The heterogeneity of MSCs can result in variations of their organic habits and remedy outcomes. Whereas large-scale MSC expansions are required to acquire a therapeutically related variety of cells for implantation, this course of can introduce cell heterogeneity. Due to this fact, improved processes are important to cut back cell heterogeneity whereas growing donor cell numbers with improved chondrogenic potential — the power of MSCs to distinguish into cartilage cells to restore cartilage tissue — to pave the best way for simpler and constant MSC-based therapies.

In a paper titled “Metabolic modulation to enhance MSC enlargement and therapeutic potential for articular cartilage restore”, printed within the scientific journal Stem Cell Analysis & Remedy, CAMP researchers detailed their growth of a priming technique to boost the enlargement of high quality MSCs by modifying the best way cells make the most of vitality. The analysis findings have proven a constructive correlation between chondrogenic potential and oxidative phosphorylation (OXPHOS), a course of that harnesses the discount of oxygen to create adenosine triphosphate — a supply of vitality that drives and helps many processes in dwelling cells. This means that manipulating MSC metabolism is a promising technique for enhancing chondrogenic potential.

Utilizing novel PATs developed by CAMP, the researchers explored the potential of metabolic modulation in each short- and long-term harvesting and reseeding of cells. To boost their chondrogenic potential, they various the nutrient composition, together with glucose, pyruvate, glutamine, and ascorbic acid (AA). As AA is reported to assist OXPHOS and its constructive affect on chondrogenic potential throughout differentiation – a course of by which immature cells turn into mature cells with particular capabilities, the researchers additional investigated its results throughout MSC enlargement.

The addition of AA to cell cultures for one passage throughout MSC enlargement and previous to initiation of differentiation was discovered to enhance chondrogenic differentiation, which is a crucial high quality attribute (CQA) for higher articular cartilage restore. Longer-term AA remedy led to a greater than 300-fold improve within the yield of MSCs with enhanced chondrogenic potential, diminished cell heterogeneity and cell senescence — a course of by which a cell ages and completely stops dividing however doesn’t die — when in comparison with untreated cells. AA-treated MSCs with improved chondrogenic potential confirmed a sturdy shift in metabolic profile to OXPHOS. This metabolic change correlated with μMRR measurements, which helps determine novel CQAs that may very well be applied in MSC manufacturing for articular cartilage restore.

The analysis additionally demonstrates the potential of the method analytical software (PAT) developed by CAMP, micromagnetic resonance relaxometry (μMRR) – a miniature benchtop gadget that employs magnetic resonance imaging (MRI) imaging on a microscopic scale – as a process-monitoring software for the enlargement of MSCs with AA supplementation. Initially used as a label-free malaria prognosis methodology as a result of presence of paramagnetic hemozoin particles, μMRR was used within the analysis to detect senescence in MSCs. This speedy, label-free methodology requires solely a small variety of cells for analysis, which permits for MSC remedy manufacturing in closed techniques — a system for shielding pharmaceutical merchandise by decreasing contamination dangers from the exterior atmosphere — whereas enabling intermittent monitoring of a restricted lot measurement per manufacturing.

“Donor-to-donor variation, intrapopulation heterogeneity and mobile senescence have impeded the success of MSCs as a regular of care remedy for articular cartilage restore. Our analysis confirmed that AA supplementation throughout MSC enlargement can overcome these bottlenecks and improve MSC chondrogenic potential,” stated Ching Ann Tee, Senior Postdoctoral Affiliate at SMART CAMP, and first creator of the paper.

By controlling metabolic situations resembling AA supplementation, coupled with CAMP’s course of analytical instruments resembling µMRR, the yield and high quality of cell remedy merchandise may very well be considerably elevated. This breakthrough might assist make MSC remedy a simpler and viable remedy possibility and supply requirements for bettering the manufacturing pipeline.”

Ching Ann Tee, Senior Postdoctoral Affiliate, SMART CAMP

“This method of using metabolic modulation to enhance MSC chondrogenic potential may very well be tailored into comparable ideas for different therapeutic indications resembling osteogenic potential for bone restore or different sorts of stem cells. Implementing our findings in MSC manufacturing settings may very well be a big step ahead for sufferers with osteoarthritis and different joint illnesses, as we are able to effectively produce giant portions of high-quality MSCs with constant performance and allow the remedy of extra sufferers,” added Prof Laurie A. Boyer, Principal Investigator at SMART CAMP, Professor of Biology and Organic Engineering at MIT and corresponding creator of the paper.

The analysis is carried out by SMART and supported by the Nationwide Analysis Basis (NRF) Singapore underneath its Campus for Analysis Excellence and Technological Enterprise (CREATE) programme.

Supply:

Singapore-MIT Alliance for Analysis and Know-how (SMART)

Journal reference:

Tee, C.A., et al. (2024) Metabolic modulation to enhance MSC enlargement and therapeutic potential for articular cartilage restore. Stem Cell Analysis & Remedy. doi.org/10.1186/s13287-024-03923-w.



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Tags: articularboostscartilageefficacyMethodMSCrepair
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