scholarly journals Protocols for Culturing and Imaging a Human Ex Vivo Osteochondral Model for Cartilage Biomanufacturing Applications

Materials ◽  
2019 ◽  
Vol 12 (4) ◽  
pp. 640 ◽  
Author(s):  
Serena Duchi ◽  
Stephanie Doyle ◽  
Timon Eekel ◽  
Cathal D. O’Connell ◽  
Cheryl Augustine ◽  
...  
Keyword(s):  
Ex Vivo ◽  
Cartilage Regeneration ◽  
Light Sheet ◽  
Cartilage Tissue ◽  
Cartilage Defects ◽  
Label Free ◽  
Microscopy Imaging ◽  
Microscopy Technique

Cartilage defects and diseases remain major clinical issues in orthopaedics. Biomanufacturing is now a tangible option for the delivery of bioscaffolds capable of regenerating the deficient cartilage tissue. However, several limitations of in vitro and experimental animal models pose serious challenges to the translation of preclinical findings into clinical practice. Ex vivo models are of great value for translating in vitro tissue engineered approaches into clinically relevant conditions. Our aim is to obtain a viable human osteochondral (OC) model to test hydrogel-based materials for cartilage repair. Here we describe a detailed step-by-step framework for the generation of human OC plugs, their culture in a perfusion device and the processing procedures for histological and advanced microscopy imaging. Our ex vivo OC model fulfils the following requirements: the model is metabolically stable for a relevant culture period of 4 weeks in a perfusion bioreactor, the processing procedures allowed for the analysis of 3 different tissues or materials (cartilage, bone and hydrogel) without compromising their integrity. We determined a protocol and the settings for a non-linear microscopy technique on label free sections. Furthermore, we established a clearing protocol to perform light sheet-based observations on the cartilage layer without the need for tedious and destructive histological procedures. Finally, we showed that our OC system is a clinically relevant in terms of cartilage regeneration potential. In conclusion, this OC model represents a valuable preclinical ex vivo tool for studying cartilage therapies, such as hydrogel-based bioscaffolds, and we envision it will reduce the number of animals needed for in vivo testing.

scholarly journals Spatiotemporal regulation of endogenous MSCs using a functional injectable hydrogel system for cartilage regeneration

2021 ◽  
Vol 13 (1) ◽  
Author(s):  
Yunsheng Dong ◽  
Yufei Liu ◽  
Yuehua Chen ◽  
Xun Sun ◽  
Lin Zhang ◽  
...  
Keyword(s):  
Cartilage Tissue Engineering ◽  
Cartilage Regeneration ◽  
Cartilage Tissue ◽  
Cartilage Defects ◽  
In Vivo Experiments ◽  
Spatiotemporal Regulation ◽  
Stromal Cell Derived Factor ◽  
Hydrogel System

AbstractHydrogels have been extensively favored as drug and cell carriers for the repair of knee cartilage defects. Recruiting mesenchymal stem cells (MSCs) in situ to the defect region could reduce the risk of contamination during cell delivery, which is a highly promising strategy to enhance cartilage repair. Here, a cell-free cartilage tissue engineering (TE) system was developed by applying an injectable chitosan/silk fibroin hydrogel. The hydrogel system could release first stromal cell-derived factor-1 (SDF-1) and then kartogenin (KGN) in a unique sequential drug release mode, which could spatiotemporally promote the recruitment and chondrogenic differentiation of MSCs. This system showed good performance when formulated with SDF-1 (200 ng/mL) and PLGA microspheres loaded with KGN (10 μΜ). The results showed that the hydrogel had good injectability and a reticular porous structure. The microspheres were distributed uniformly in the hydrogel and permitted the sequential release of SDF-1 and KGN. The results of in vitro experiments showed that the hydrogel system had good cytocompatibility and promoted the migration and differentiation of MSCs into chondrocytes. In vivo experiments on articular cartilage defects in rabbits showed that the cell-free hydrogel system was beneficial for cartilage regeneration. Therefore, the composite hydrogel system shows potential for application in cell-free cartilage TE.

scholarly journals Regenerative Medicine: where their origin makes species meet

2019 ◽  
Vol 37 (3) ◽  
pp. 6-7
Author(s):  
Jos Malda
Keyword(s):  
Articular Cartilage ◽  
Ex Vivo ◽  
Mammalian Species ◽  
Cartilage Tissue ◽  
In Vitro Assays ◽  
Cartilage Defects ◽  
In Vivo Models ◽  
Societal Pressure

The articular cartilage of joints serves diverse functions, including absorbing shock, transmitting force, and enabling low-friction joint motion. Regeneration of articular cartilage defects remains, however, a significant challenge in both human and veterinary orthopaedic practice. Ex vivo and in vivo models play a crucial role in translating novel potential regenerative treatments from bench to bedside. However, in view of the predictive power of these models and the One Medicine concept that proclaim that there should be no dividing lines between human and animal medicine to learn, it is important to understand the similarities as well as differences in the cartilage tissue between species. To this aim, osteochondral cores of the femoral condyles were studied in 58 different mammalian species ranging from mouse to elephant. Interestingly, while biochemical composition remained relatively constant, cartilage thickness and cellularity were similar underscoring the importance of the equine species as a model for human orthopaedic interventions. Nevertheless, political ambition and societal pressure are now asking for a drastically reduction of animal experimentation and have further spiked the development of more predictive in vitro and ex vivo models. In addition to a range of more sophisticated in vitro assays, this has now also provided an ex vivo osteochondral defect model that can be generated based on equine donor tissue. Although such models better represent the situation in the native tissue and can be used to assess (osteo)chondral repair strategies, they still lack some important aspects of the in vivo (patho)physiological (inflammatory) environment, as well as the exposure to mechanical loading. This illustrates the challenges that we still face in translating these novel approaches from bench to bedside.

Combined Effects of Bone Marrow-Derived Mesenchymal Stem Cells and PLGA-PEG-PLGA Scaffolds on Repair of Articular Cartilage Defect

2013 ◽  
Vol 815 ◽  
pp. 345-349 ◽  
Author(s):  
Ching Wen Hsu ◽  
Ping Liu ◽  
Song Song Zhu ◽  
Feng Deng ◽  
Bi Zhang
Keyword(s):  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
Articular Cartilage ◽  
Growth Factor ◽  
Cartilage Defect ◽  
Cartilage Regeneration ◽  
Tissue Growth ◽  
Cartilage Defects

Here we reported a combined technique for articular cartilage repair, consisting of bone arrow mesenchymal stem cells (BMMSCs) and poly (dl-lactide-co-glycolide-b-ethylene glycol-b-dl-lactide-co-glycolide) (PLGA-PEG-PLGA) triblock copolymers carried with tissue growth factor (TGF-belat1). In the present study, BMMSCs seeded on PLGA-PEG-PLGA with were incubated in vitro, carried or not TGF-belta1, Then the effects of the composite on repair of cartilage defect were evaluated in rabbit knee joints in vivo. Full-thickness cartilage defects (diameter: 5 mm; depth: 3 mm) in the patellar groove were either left empty (n=18), implanted with BMMSCs/PLGA (n=18), TGF-belta1 modified BMMSCs/PLGA-PEG-PLGA. The defect area was examined grossly, histologically at 6, 24 weeks postoperatively. After implantation, the BMMSCs /PLGA-PEG-PLGA with TGF-belta1 group showed successful hyaline-like cartilage regeneration similar to normal cartilage, which was superior to the other groups using gross examination, qualitative and quantitative histology. These findings suggested that a combination of BMMSCs/PLGA-PEG-PLGA carried with tissue growth factor (TGF-belat1) may be an alternative treatment for large osteochondral defects in high loading sites.

scholarly journals Cartilage Extracellular Matrix Scaffold With Kartogenin-Encapsulated PLGA Microspheres for Cartilage Regeneration

2020 ◽  
Vol 8 ◽  
Author(s):  
Yanhong Zhao ◽  
Xige Zhao ◽  
Rui Zhang ◽  
Ying Huang ◽  
Yunjie Li ◽  
...  
Keyword(s):  
Extracellular Matrix ◽  
Composite Scaffold ◽  
Rabbit Model ◽  
Cartilage Tissue ◽  
Cartilage Defects ◽  
Specific Marker ◽  
Molecular Compound ◽  
Plga Microspheres

Repair of articular cartilage defects is a challenging aspect of clinical treatment. Kartogenin (KGN), a small molecular compound, can induce the differentiation of bone marrow-derived mesenchymal stem cells (BMSCs) into chondrocytes. Here, we constructed a scaffold based on chondrocyte extracellular matrix (CECM) and poly(lactic-co-glycolic acid) (PLGA) microspheres (MP), which can slowly release KGN, thus enhancing its efficiency. Cell adhesion, live/dead staining, and CCK-8 results indicated that the PLGA(KGN)/CECM scaffold exhibited good biocompatibility. Histological staining and quantitative analysis demonstrated the ability of the PLGA(KGN)/CECM composite scaffold to promote the differentiation of BMSCs. Macroscopic observations, histological tests, and specific marker analysis showed that the regenerated tissues possessed characteristics similar to those of normal hyaline cartilage in a rabbit model. Use of the PLGA(KGN)/CECM scaffold may mimic the regenerative microenvironment, thereby promoting chondrogenic differentiation of BMSCs in vitro and in vivo. Therefore, this innovative composite scaffold may represent a promising approach for acellular cartilage tissue engineering.

scholarly journals Progress and Applications of Polyphosphate in Bone and Cartilage Regeneration

2019 ◽  
Vol 2019 ◽  
pp. 1-12 ◽  
Author(s):  
Yan Wang ◽  
Min Li ◽  
Pei Li ◽  
Haijun Teng ◽  
Dehong Fan ◽  
...  
Keyword(s):  
Cartilage Regeneration ◽  
High Energy ◽  
Cartilage Defects ◽  
Artificial Bone ◽  
Cellular Mechanisms ◽  
Inorganic Polyphosphate ◽  
Morphogenetic Effects ◽  
And Cartilage

Patients with bone and cartilage defects due to infection, tumors, and trauma are quite common. Repairing bone and cartilage defects is thus a major problem for clinicians. Autologous and artificial bone transplantations are associated with many challenges, such as limited materials and immune rejection. Bone and cartilage regeneration has become a popular research topic. Inorganic polyphosphate (polyP) is a widely occurring biopolymer with high-energy phosphoanhydride bonds that exists in organisms from bacteria to mammals. Much data indicate that polyP acts as a regulator of gene expression in bone and cartilage tissues and exerts morphogenetic effects on cells involved in bone and cartilage formation. Exposure of these cells to polyP leads to the increase of cytokines that promote the differentiation of mesenchymal stem cells into osteoblasts, accelerates the osteoblast mineralization process, and inhibits the differentiation of osteoclast precursors to functionally active osteoclasts. PolyP-based materials have been widely reported in in vivo and in vitro studies. This paper reviews the current cellular mechanisms and material applications of polyP in bone and cartilage regeneration.

scholarly journals Investigation of molecular parameters on cartilage regeneration in experimental models of cartilage defects and osteoarthritis

2014 ◽  
Author(s):  
Ελευθέριος Μακρής
Keyword(s):  
Cartilage Regeneration ◽  
Experimental Models ◽  
Cartilage Defects ◽  
Molecular Parameters

Ο αρθρικός και ο ινώδης χόνδρος έχουν περιορισμένη ικανότητα αναγέννησης μετά από τραυματικές κακώσεις και παθήσεις των αρθρώσεων. Δεδομένου του κριτικού ρόλου των ιστών αυτών στην προστασία των αρθρικών οστικών δομών και στην εξασφάλιση σταθερών λειτουργικών αρθρώσεων, η ανάπτυξη μεθόδων που προάγουν την αναγέννηση ή / και την επιδιορθώση των ιστών αυτών ειναι εξαιρετικά σημαντική. Η επιστήμη της ανάπτυξης μηχανικών ιστών παρέχει σήμερα εξαιρετικές προοπτικές στη θεραπεία των εκφυλισμένων και παθολογικών χόνδρινων ιστών. Και ενώ μεγάλη πρόοδος έχει επιτευχθεί πρόσφατα ως προς την ανάπτυξη νεοχόνδρινων ιστών με θλιπτικές μηχανικές ιδιότητες ανάλογες των φυσικών ιστών, ωστόσο οι ιδιότητες εφελκυσμού των νεοϊστών ειναι συγκριτικά υποδιαίστερες. Ως εκ τούτου, η ανάγκη ανάπτυξης νέων βελτιωμένων μεθόδων για να δημιουργία λειτουργικών νεοϊστών και την αποτελεσματική αντιμετώπιση των περίπου 46.4 εκατομμυρίων ασθένών που υποφέρουν σήμερα από αρθρίτιδα μόνο στις Ηνωμένες Πολιτείες, καθίσταται ιδιαίτερα επιτακτική.Δεδομένης της προόδου που έχει επιτευχθεί την τελευταία δεκαετία στην ανάπτυξη μηχανικών χόνδρινων ιστών, η παρούσα διδακτορική διατριβή έχει τρεις γενικούς στόχους: 1) τη διερεύνηση νέων μοριακών/βιολογικών μεθόδων αναγέννησης του αρθρικού χόνδρου σε in vitro και in vivo πειραματικά μοντέλα χόνδρινων βλαβών και οστεοαρθρίτιδας, 2) την ανάπτυξη ή/και βελτίωση μεθόδων αναγέννησης νεοχόνδρινών ιστών με ευρεία κλινική εφαρμογή, και 3) την ανάπτυξη μεθόδων βιολογικής και μηχανικής ωρίμανσης των νεοϊστών και μεθόδων ενσωμάτωσης τους με το φυσικό ιστό μετά απο μεταμόσχευση. Για την επίτευξη των στόχων αυτών, η παρούσα διατριβή περιγράφει την διερεύνηση νέων εξωγενών βιολογικών παραγόντων για την ανάπτυξη μηχανικών νεοϊστών με δομική οργάνωση ανάλογης των αντίστοιχων φυσικών ιστών. Αρχικά, μελετήθηκε η χρήση του χαλκού και της υδροξυλυσίνης, δύο σημαντικών μεσολαβητών της φυσιολογικής διαδικασίας ανάπτυξης διασταυρώμενων δέσμών κολλαγόνου, για την ενίσχυση της περιεκτικότητας των διασταυρώμενων δεσμών κολλαγόνου στους μηχανικούς ιστούς. Αναλόγως, σε μια διαφορετική μελέτη διερευνήθηκε η in vitro καλλιέργεια φυσικών μυοσκελετικών ιστών και μηχανικών νεοϊστών σε υποξικό περιβάλλον, με στόχο την ενίσχυση της περιεκτικότητάς τους σε διασταυρώμενους δεσμούς κολλαγόνου, και ως εκ τούτου, τη βελτίωση των λειτουργικών τους ιδιοτήτων. Λαμβάνοντας υπόψη την εκταινώς περιγεγραμένη στη βιβλιογραφία θετική συσχέτιση μεταξύ της περιεκτικότητας διασταυρούμενων δεσμών κολλαγόνου και εφελκυστικών μηχανικών ιδιοτήτων των νεοϊστών, μελετήθηκε μία νέα μέθοδος για την αύξηση αυτών των διασταυρούμενων δεσμών κολλαγόνου βασιζόμενη στην εξωγενή χορήγηση του ενζύμου λυσυλική οξειδάση (LOX) κατά τη καλλιέργεια των νεοϊστών, με απώτερο στόχο την περαιτέρω ενισχυση των λειτουργικών τους ιδιοτήτων.Άλλες μελέτες της παρούσας διατριβής επικεντρώνονται στην ανάπτυξη μεθόδων ενίσχυσης της περιεκτικότητας των μηχανικών ιστών σε κολλαγόνο, που όταν συνδυάζονται με τις τεχνικές ενίσχυσης των διασταυρούμενων δεσμών κολλαγόνου θα μπορούσαν να βελτιώσουν περαιτερώ την μηχανική ωρίμανση των νεοϊστών για την αποτελεσματική αποκατάσταση χόνδρινών ελλειμάτων σε πειραματικά μοντέλα αρθρικών παθήσεων. Ειδικότερα, δύο παράγοντες που είναι γνωστοί ρυθμιστές της ενδοκυτταρικής σηματοδότησης του Ca2+ (η διγοξίνη και η τριφωσφορική αδενοσίνη), εξετάστηκαν ως εναλλακτικές μέθοδοι για τη μηχανική βελτίωση των ιστών μέσω αύξησης της περιεκτικότητάς τους σε κολλαγόνο και σε διασταυρούμενους δεσμούς κολλαγόνου. Επιπλέον, ανάλογες μελέτες επικεντρώθηκαν στην βελτίωση μιας θεραπευτικής αγωγής που περιλαμβάνει το βιοφυσικό παράγοντα χονδροϊτινάση-ABC (C-ABC) και τον βιοχημικό παράγοντα αυξητικό παράγοντα μετασχηματισμού -β1 (ΤGF-β1) με στόχο πάλι την ενισχύση τών λειτουργικών ιδιοτήτων των νεοχόνδρινων ιστών. Μια διαφορετική μελέτη εξέτασε την ικανότητα της εμβιομηχανικής διέγερσης με τη μορφή παθητικής αξονικής θλιπτικής φόρτισης να συμβάλλει σε in vitro ανάπτυξη νεοχόνδρινών ιστών με προδιαγεγραμμένο σχήμα/αρχιτεκτονική. Τέλος, μια πρόσθετη μελέτη διερεύνησε την χρήση χονδροκυττάρων απο την ποδοκνημική άρθρωση για την κατασκευή νεοχόνδρινων μοσχευμάτων προοριζόμενων για την βλάβες της ποδοκνημικής άρθρωσης.Καταλεικτικά, η παρούσα διατριβή διερεύνησε τη δυνατότητα ενσωμάτωσης του νεοχόνδρινου με τον φυσικό ιστό μέσω της εξωγενούς χορήγησης του ενζύμου LOX. Μια αρχική μελέτη διερεύνησε τη δυνατότητα του LOX να προώθηση της ενσωμάτωσης μεταξύ του νεοχόνδρου και του φυσικού αρθρικού χόνδρου σε in vitro περιβάλλον καλλιέργειας των ιστών. Ελπιδοφόρα αποτελέσματα αυτής της μέλετης οδήγησαν σε συνδυασμό του ενζύμου LOX με τους παράγοντες C-ABC και ΤGF-β1 με στόχο την ταυτόχρονη ενίσχυση της ωρίμανσης του νεοχόνδρου και τη ενσωμάτωση του με τον φυσικό αρθρικό χόνδρο. Ειδικότερα, ο συνδυασμός των παραγόντων αυτών εφαρμόστηκε πρώτα σε ένα in vitro μοντέλο ενσωμάτωσης των δύο ιστών, και στη συνέχεια σε ένα πειραματικό ζωικό μοντέλο για τη διερεύνηση της ικανότητας του in νίνο περιβάλλοντος να ενισχύσει περαιτέρω της βιολογική αυτή διεργασία.Σε γενικές γραμμές, η σειρά των μελετών που περιγράφονται στην παρούσα διατριβή αντιπροσωπεύουν πρωτοπόρες και δυνητικώς κλινικά εφαρμόσιμες μοριακές μεθόδους για την ενίσχυση των λειτουργικών ιδιοτήτων ενός μεγάλου φάσματος κολλαγόνων ιστών και νεοϊστών. Συγκεκριμένα, ο χαλκός, η ενδογενής παραγώγη του ενζύμου LOX μέσω της καλλιεργειας των ιστών σε υποξίκό περιβάλλον, και η εξωγενή χορήγηση του LOX βελτιώνουν τις μηχανικές ιδιότητες των μηχανικών ιστών αναδεικνύοντας την ικανότητα στόχευσης της αύξησης των διαστραυρούμενων δεσμών κολλαγόνου για την ανάπτυξη βιομιμητικών και μηχανικά ισχυρών νεοχονδρινών ιστών. Εναλλακτικά, η διγοξίνη, η τριφωσφορική αδενοσίνη, ο C-ABC, ο ΤGF -β1, και η στατική εφαρμογή βάρους αποτελουν αποτελεσματικές μεθόδους για την ενίσχυση του κολλαγόνου και την αυξηση των διαστραυρούμενων δεσμών κολλαγόνου στους νεοϊστούς. Τέλος, η εξωγενής χορήγηση του LOX, μονομερώς ή σε συνδυασμό με τους παράγοντες C-ABC και ΤGF -β1, βρέθηκε να προάγει την in vitro και in vivo ωρίμανση των νεοϊστών και την ενσωμάτωση τους με το φυσικό ιστό, αντανακλώντας την πολλά υπόσχόμενη συνδιαστική χρήση των παραγόντων αυτών για την θεραπευτική αντιμετώπιση χόνδρινων βλαβών in νίνο. Συνολικά, οι μελέτες που περιγράφονται στην παρούσα διατριβή αποτελούν μια επισκόπηση διαφορετικών και πολλά υποσχόμενων νέων τεχνολογιών με στόχο 1) την ενίσχυση των μηχανικών ιδιοτήτων των φυσικών ιστών και νεοϊστών, και 2) την ενίσχυση των δυνατοτήτων τους να ενσωματώνονται in vitro, υπογραμμίζοντας τη δυνατότητα για εφαρμογή των τεχνολογιών αυτών στη ανάπτυξη κλινικά λειτουργικών νεοϊστών για την επιδιόρθωση και / ή αντικατάσταση των ιστών που έχουν καταστραφεί από τραυματισμό ή ασθένεια.

scholarly journals Chondrogenic Potential of Dental-Derived Mesenchymal Stromal Cells

Osteology ◽  
2021 ◽  
Vol 1 (3) ◽  
pp. 149-174
Author(s):  
Naveen Jeyaraman ◽  
Gollahalli Shivashankar Prajwal ◽  
Madhan Jeyaraman ◽  
Sathish Muthu ◽  
Manish Khanna
Keyword(s):  
Tissue Engineering ◽  
Mesenchymal Stromal Cells ◽  
Tissue Regeneration ◽  
Stromal Cells ◽  
Cartilage Tissue Engineering ◽  
Cartilage Regeneration ◽  
Cartilage Tissue ◽  
Dentin Matrix

The field of tissue engineering has revolutionized the world in organ and tissue regeneration. With the robust research among regenerative medicine experts and researchers, the plausibility of regenerating cartilage has come into the limelight. For cartilage tissue engineering, orthopedic surgeons and orthobiologists use the mesenchymal stromal cells (MSCs) of various origins along with the cytokines, growth factors, and scaffolds. The least utilized MSCs are of dental origin, which are the richest sources of stromal and progenitor cells. There is a paradigm shift towards the utilization of dental source MSCs in chondrogenesis and cartilage regeneration. Dental-derived MSCs possess similar phenotypes and genotypes like other sources of MSCs along with specific markers such as dentin matrix acidic phosphoprotein (DMP) -1, dentin sialophosphoprotein (DSPP), alkaline phosphatase (ALP), osteopontin (OPN), bone sialoprotein (BSP), and STRO-1. Concerning chondrogenicity, there is literature with marginal use of dental-derived MSCs. Various studies provide evidence for in-vitro and in-vivo chondrogenesis by dental-derived MSCs. With such evidence, clinical trials must be taken up to support or refute the evidence for regenerating cartilage tissues by dental-derived MSCs. This article highlights the significance of dental-derived MSCs for cartilage tissue regeneration.

scholarly journals 3D Cartilage Regeneration With Certain Shape and Mechanical Strength Based on Engineered Cartilage Gel and Decalcified Bone Matrix

2021 ◽  
Vol 9 ◽  
Author(s):  
Zheng Ci ◽  
Ying Zhang ◽  
Yahui Wang ◽  
Gaoyang Wu ◽  
Mengjie Hou ◽  
...  
Keyword(s):  
Mechanical Strength ◽  
Bone Matrix ◽  
Cartilage Regeneration ◽  
Cartilage Tissue ◽  
Cartilage Defects ◽  
High Porosity ◽  
Good Biocompatibility ◽  
Engineered Cartilage ◽  
Decalcified Bone Matrix

Scaffold-free cartilage-sheet technology can stably regenerate high-quality cartilage tissue in vivo. However, uncontrolled shape maintenance and mechanical strength greatly hinder its clinical translation. Decalcified bone matrix (DBM) has high porosity, a suitable pore structure, and good biocompatibility, as well as controlled shape and mechanical strength. In this study, cartilage sheet was prepared into engineered cartilage gel (ECG) and combined with DBM to explore the feasibility of regenerating 3D cartilage with controlled shape and mechanical strength. The results indicated that ECG cultured in vitro for 3 days (3 d) and 15 days (15 d) showed good biocompatibility with DBM, and the ECG–DBM constructs successfully regenerated viable 3D cartilage with typical mature cartilage features in both nude mice and autologous goats. Additionally, the regenerated cartilage had comparable mechanical properties to native cartilage and maintained its original shape. To further determine the optimal seeding parameters for ECG, the 3 d ECG regenerated using human chondrocytes was diluted in different concentrations (1:3, 1:2, and 1:1) for seeding and in vivo implantation. The results showed that the regenerated cartilage in the 1:2 group exhibited better shape maintenance and homogeneity than the other groups. The current study established a novel mode of 3D cartilage regeneration based on the design concept of steel (DBM)-reinforced concrete (ECG) and successfully regenerated homogenous and mature 3D cartilage with controlled shape and mechanical strength, which hopefully provides an ideal cartilage graft for the repair of various cartilage defects.

scholarly journals Effect of different aged cartilage ECM on chondrogenesis of BMSCs in vitro and in vivo

2020 ◽  
Vol 7 (6) ◽  
pp. 583-595
Author(s):  
Xiuyu Wang ◽  
Yan Lu ◽  
Wan Wang ◽  
Qiguang Wang ◽  
Jie Liang ◽  
...  
Keyword(s):  
Articular Cartilage ◽  
Current Knowledge ◽  
Cartilage Tissue Engineering ◽  
Cartilage Regeneration ◽  
Biological Properties ◽  
Cartilage Tissue ◽  
Before And After ◽  
Marrow Mesenchymal Stem Cells

Abstract Extracellular matrix (ECM)-based biomaterials are promising candidates in cartilage tissue engineering by simulating the native microenvironment to regulate the chondrogenic differentiation of bone marrow mesenchymal stem cells (BMSCs) without exogenous growth factors. The biological properties of ECM scaffolds are primarily depended on the original source, which would directly influence the chondrogenic effects of the ECM materials. Despite the expanding investigations on ECM scaffolds in recent years, the selection of optimized ECM materials in cartilage regeneration was less reported. In this study, we harvested and compared the articular cartilage ECM from newborn, juvenile and adult rabbits. The results demonstrated the significant differences in the mechanical strength, sulphated glycosaminoglycan and collagen contents of the different aged ECM, before and after decellularization. Consequently, different compositional and mechanical properties were shown in the three ECM-based collagen hydrogels, which exerted age-dependent chondrogenic inducibility. In general, both in vitro and in vivo results suggested that the newborn ECM promoted the most chondrogenesis of BMSCs but led to severe matrix calcification. In contrast, BMSCs synthesized the lowest amount of cartilaginous matrix with minimal calcification with adult ECM. The juvenile ECM achieved the best overall results in promoting chondrogenesis of BMSCs and preventing matrix calcification. Together, this study provides important information to our current knowledge in the design of future ECM-based biomaterials towards a successful repair of articular cartilage.

scholarly journals A Single-Cell Culture System for Dissecting Microenvironmental Signaling in Development and Disease of Cartilage Tissue

2021 ◽  
Vol 9 ◽  
Author(s):  
Jade Tassey ◽  
Arijita Sarkar ◽  
Ben Van Handel ◽  
Jinxiu Lu ◽  
Siyoung Lee ◽  
...  
Keyword(s):  
Single Cell ◽  
Articular Chondrocytes ◽  
3D Culture ◽  
Cartilage Regeneration ◽  
Culture Method ◽  
Cartilage Tissue ◽  
Surrounding Matrix ◽  
Clonogenic Potential

Cartilage tissue is comprised of extracellular matrix and chondrocytes, a cell type with very low cellular turnover in adults, providing limited capacity for regeneration. However, in development a significant number of chondrocytes actively proliferate and remodel the surrounding matrix. Uncoupling the microenvironmental influences that determine the balance between clonogenic potential and terminal differentiation of these cells is essential for the development of novel approaches for cartilage regeneration. Unfortunately, most of the existing methods are not applicable for the analysis of functional properties of chondrocytes at a single cell resolution. Here we demonstrate that a novel 3D culture method provides a long-term and permissive in vitro niche that selects for highly clonogenic, colony-forming chondrocytes which maintain cartilage-specific matrix production, thus recapitulating the in vivo niche. As a proof of concept, clonogenicity of Sox9IRES–EGFP mouse chondrocytes is almost exclusively found in the highest GFP+ fraction known to be enriched for chondrocyte progenitor cells. Although clonogenic chondrocytes are very rare in adult cartilage, we have optimized this system to support large, single cell-derived chondrogenic organoids with complex zonal architecture and robust chondrogenic phenotype from adult pig and human articular chondrocytes. Moreover, we have demonstrated that growth trajectory and matrix biosynthesis in these organoids respond to a pro-inflammatory environment. This culture method offers a robust, defined and controllable system that can be further used to interrogate the effects of various microenvironmental signals on chondrocytes, providing a high throughput platform to assess genetic and environmental factors in development and disease.

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