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Polymers2021; 13(5); 747; doi: 10.3390/polym13050747

Equine Tenocyte Seeding on Gelatin Hydrogels Improves Elongated Morphology.

Abstract: (1) Background: Tendinopathy is a common injury in both human and equine athletes. Representative in vitro models are mandatory to facilitate translation of fundamental research into successful clinical treatments. Natural biomaterials like gelatin provide favorable cell binding characteristics and are easily modifiable. In this study, methacrylated gelatin (gel-MA) and norbornene-functionalized gelatin (gel-NB), crosslinked with 1,4-dithiotreitol (DTT) or thiolated gelatin (gel-SH) were compared. (2) Methods: The physicochemical properties (H-NMR spectroscopy, gel fraction, swelling ratio, and storage modulus) and equine tenocyte characteristics (proliferation, viability, and morphology) of four different hydrogels (gel-MA, gel-NB85/DTT, gel-NB55/DTT, and gel-NB85/SH75) were evaluated. Cellular functionality was analyzed using fluorescence microscopy (viability assay and focal adhesion staining). (3) Results: The thiol-ene based hydrogels showed a significantly lower gel fraction/storage modulus and a higher swelling ratio compared to gel-MA. Significantly less tenocytes were observed on gel-MA discs at 14 days compared to gel-NB85/DTT, gel-NB55/DTT and gel-NB85/SH75. At 7 and 14 days, the characteristic elongated morphology of tenocytes was significantly more pronounced on gel-NB85/DTT and gel-NB55/DTT in contrast to TCP and gel-MA. (4) Conclusions: Thiol-ene crosslinked gelatins exploiting DTT as a crosslinker are the preferred biomaterials to support the culture of tenocytes. Follow-up experiments will evaluate these biomaterials in more complex models.
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Publication Date: 2021-02-28 PubMed ID: 33670848PubMed Central: PMC7957613DOI: 10.3390/polym13050747Google Scholar: Lookup
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Summary

This research summary has been generated with artificial intelligence and may contain errors and omissions. Refer to the original study to confirm details provided. Submit correction.

This research investigates how seeding equine tenocytes, the cells responsible for the health and repair of tendons, on different types of gelatin hydrogels impacts their shape and function. The study found that certain types of gelatin, especially those crosslinked with 1,4-dithiotreitol (DTT), were more conducive to the maintenance and growth of tenocytes, providing guidance for future development of treatments for tendon injuries in both humans and horses.

Research Background

  • The study begins by discussing the prevalence of tendon injuries, also known as tendinopathy, in both humans and horses, particularly athletes.
  • The research emphasizes the need for good representative models to test potential treatments before they’re used in the clinic.
  • Gelatins are highlighted as natural biomaterials that are easily modified and offer good cell binding properties, making them excellent candidates for in vitro (outside of a living organism) studies.

Research Methodology

  • The researchers compared four different types of gelatin hydrogels: methacrylated gelatin (gel-MA) and three variations of norbornene-functionalized gelatin (gel-NB), which were each crosslinked with either 1,4-dithiotreitol (DTT) or thiolated gelatin (gel-SH).
  • Several testing methods were used to analyze the properties of these hydrogels and how equine tenocytes interacted with them, including H-NMR spectroscopy, gel fraction, swelling ratio, and storage modulus assessments.
  • Fluorescence microscopy was used to evaluate cell functionality, including viability (how many cells remain alive) and focal adhesion staining (which reveals the points where cells adhere to the surrounding matrix).

Research Findings

  • The results showed that gelatins crosslinked with DTT resulted in a lower gel fraction/storage modulus and a higher swelling ratio than gel-MA, indicating they have different physical properties.
  • Furthermore, fewer tenocytes were found on gel-MA discs after 14 days compared to the other types of gelatin.
  • The stretched or elongated morphology of tenocytes, typical of healthy cells of this type, was significantly more pronounced on certain types of gelatin (particularly gel-NB85/DTT and gel-NB55/DTT) than on other gel samples and tissue culture polystyrene (TCP).

Conclusion and Future Directions

  • The study concluded that gelatins crosslinked with DTT are the preferred biomaterials for nurturing tenocyte culture. This is a crucial conclusion for the field as these findings can guide the development of superior treatments for tendinopathy.
  • As future work, the researchers plan to validate these results further using more complex models and experiments.

Cite This Article

APA
Meeremans M, Van Damme L, De Spiegelaere W, Van Vlierberghe S, De Schauwer C. (2021). Equine Tenocyte Seeding on Gelatin Hydrogels Improves Elongated Morphology. Polymers (Basel), 13(5), 747. https://doi.org/10.3390/polym13050747

Publication

Polymers
ISSN: 2073-4360
NlmUniqueID: 101545357
Country: Switzerland
Language: English
Volume: 13
Issue: 5
PII: 747

Researcher Affiliations

Meeremans, Marguerite
  • Comparative Physiology, Faculty of Veterinary Medicine, Ghent University, Salisburylaan 133, B-9820 Merelbeke, Belgium.
Van Damme, Lana
  • Polymer Chemistry and Biomaterials Group, Centre of Macromolecular Chemistry, Faculty of Sciences, Ghent University, Krijgslaan 281 S4-Bis, B-9000 Ghent, Belgium.
De Spiegelaere, Ward
  • Department of Morphology, Faculty of Veterinary Medicine, Ghent University, Salisburylaan 133, B-9820 Merelbeke, Belgium.
Van Vlierberghe, Sandra
  • Polymer Chemistry and Biomaterials Group, Centre of Macromolecular Chemistry, Faculty of Sciences, Ghent University, Krijgslaan 281 S4-Bis, B-9000 Ghent, Belgium.
De Schauwer, Catharina
  • Comparative Physiology, Faculty of Veterinary Medicine, Ghent University, Salisburylaan 133, B-9820 Merelbeke, Belgium.

Grant Funding

  • BOFSTG2019004801 / Bijzonder Onderzoeksfonds UGent
  • I003318N / Fonds Wetenschappelijk Onderzoek

Conflict of Interest Statement

The authors declare no conflict of interest.

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