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. 2023 Nov 23:11:1295626.
doi: 10.3389/fbioe.2023.1295626. eCollection 2023.

In vivo biocompatibility testing of nanoparticle-functionalized alginate-chitosan scaffolds for tissue engineering applications

Nancy G Viveros-Moreno  1 , Mario Garcia-Lorenzana  2 , Eduardo Peña-Mercado  3 , Josune García-Sanmartín  4 , Judit Narro-Íñiguez  4 , Marcela Salazar-García  5 , Sara Huerta-Yepez  6 , Concepción Sanchez-Gomez  5 , Alfredo Martínez  4 , Nohra E Beltran-Vargas  3
Affiliations

In vivo biocompatibility testing of nanoparticle-functionalized alginate-chitosan scaffolds for tissue engineering applications

Nancy G Viveros-Moreno et al. Front Bioeng Biotechnol. .

Abstract

Background: There is a strong interest in designing new scaffolds for their potential application in tissue engineering and regenerative medicine. The incorporation of functionalization molecules can lead to the enhancement of scaffold properties, resulting in variations in scaffold compatibility. Therefore, the efficacy of the therapy could be compromised by the foreign body reaction triggered after implantation. Methods: In this study, the biocompatibilities of three scaffolds made from an alginate-chitosan combination and functionalized with gold nanoparticles (AuNp) and alginate-coated gold nanoparticles (AuNp + Alg) were evaluated in a subcutaneous implantation model in Wistar rats. Scaffolds and surrounding tissue were collected at 4-, 7- and 25-day postimplantation and processed for histological analysis and quantification of the expression of genes involved in angiogenesis, macrophage profile, and proinflammatory (IL-1β and TNFα) and anti-inflammatory (IL-4 and IL-10) cytokines. Results: Histological analysis showed a characteristic foreign body response that resolved 25 days postimplantation. The intensity of the reaction assessed through capsule thickness was similar among groups. Functionalizing the device with AuNp and AuNp + Alg decreased the expression of markers associated with cell death by apoptosis and polymorphonuclear leukocyte recruitment, suggesting increased compatibility with the host tissue. Similarly, the formation of many foreign body giant cells was prevented. Finally, an increased detection of alpha smooth muscle actin was observed, showing the angiogenic properties of the elaborated scaffolds. Conclusion: Our results show that the proposed scaffolds have improved biocompatibility and exhibit promising potential as biomaterials for elaborating tissue engineering constructs.

Keywords: alginate; biocompatibility; chitosan; foreign body reaction; subcutaneous implantation.

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Conflict of interest statement

The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.

Figures

FIGURE 1
FIGURE 1
(A) Macroscopic appearance of a hydrated chitosan–alginate scaffold. (B) Subdermal implantation sites in the dorsal region, no signs of infection or rejection are appreciated. (C, D) General view of tissues collected after sacrifice.
FIGURE 2
FIGURE 2
Representative photomicrographs of subcutaneously implanted chitosan-alginate scaffolds showing FBR and the development and evolution of the capsule over time, (H–E; bar = 1 mm). Identifiers: C: capsule, LI: leukocyte infiltrate, S: scaffold. The thickness of the capsule is shown between the dashed lines. After 25 days, the capsule decreases. Framed regions were enlarged in Figure 3 to show the details of the cellular infiltrates.
FIGURE 3
FIGURE 3
(A) Characterization of the response to a foreign body, (H-E; bar = 100 µm). Identifiers: Arrow: foreign body giant cells; arrowhead: myxoid areas; N: neutrophils; star: necrosis; S: scaffold. Graphical representation of (B) cell density, (C) percentage of area occupied by the cellular infiltrate, and (D) density of foreign body giant cells. *p < 0.01, **p < 0.001. Data are presented as mean ± SEM.
FIGURE 4
FIGURE 4
Inflammatory response. (A) qRT-PCR showing the levels of proinflammatory (IL-1β and TNFα) and anti-inflammatory (IL-4 and IL-10) cytokines. GAPDH was used as a housekeeping gene. (B) Representative immunohistochemical images and quantification of anti-Iba1 on the scaffolds at different points of time, (bar = 50 μm). Data are presented as mean ± SEM. (*) p < 0.05.
FIGURE 5
FIGURE 5
Representative photomicrographs of capsule thickness and internal collagen between groups over time (A), (Masson; bar = 1 mm). Framed regions were enlarged (Masson; x 200) to show collagen fibers. Identifiers: star: capsule; white arrow: collagen fibers. (B) Graphical representation of capsule thickness. (C) Graphical representation of collagen deposits inside the scaffolds. Data are presented as mean ± SEM. **p < 0.001.
FIGURE 6
FIGURE 6
Macrophage profile. qRT-PCR values showing the expression of markers for the M1 (TNFα, CD11, TLR4, INOS, and CD86) and M2 phenotype (IL4, IL10, VEGF, Mrc1, and ARG1) in all scaffolds. Data are presented as mean ± SEM.
FIGURE 7
FIGURE 7
Angiogenic response. (A) qRT-PCR showing the levels of PECAM1 and VEGF, (B) Representative immunohistochemical images and quantification analysis of anti-αSMA on the scaffolds at different points of time, (bar = 50 μm). (*) p < 0.05, (**) p < 0.01, (***) p < 0.001. Data are presented as mean ± SEM.
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References

    1. Abaricia J. O., Farzad N., Heath T. J., Simmons J., Morandini L., Olivares-Navarrete R. (2021). Control of innate immune response by biomaterial surface topography, energy, and stiffness. Acta Biomater. 133, 58–73. 10.1016/j.actbio.2021.04.021 - DOI - PMC - PubMed
    1. Anderson J. M., Rodriguez A., Chang D. T. (2008). Foreign body reaction to biomaterials. Semin. Immunol. 20 (2), 86–100. 10.1016/j.smim.2007.11.004 - DOI - PMC - PubMed
    1. Baranes K., Shevach M., Shefi O., Dvir T. (2016). Gold nanoparticle-decorated scaffolds promote neuronal differentiation and maturation. Nano Lett. 16 (5), 2916–2920. 10.1021/acs.nanolett.5b04033 - DOI - PubMed
    1. Barone D. G., Carnicer-Lombarte A., Tourlomousis P., Hamilton R. S., Prater M., Rutz A. L., et al. (2022). Prevention of the foreign body response to implantable medical devices by inflammasome inhibition. Proc. Natl. Acad. Sci. U. S. A. 119 (12), e2115857119. 10.1073/pnas.2115857119 - DOI - PMC - PubMed
    1. Beltran-Vargas N. E., Peña-Mercado E., Sánchez-Gómez C., Garcia-Lorenzana M., Ruiz J. C., Arroyo-Maya I., et al. (2022). Sodium alginate/chitosan scaffolds for cardiac tissue engineering: the influence of its three-dimensional material preparation and the use of gold nanoparticles. Polym. (Basel) 14 (16), 3233. 10.3390/polym14163233 - DOI - PMC - PubMed

Grants and funding

The authors declare financial support was received for the research, authorship, and/or publication of this article. Children’s Hospital of Mexico Federico Gomez (protocols HIM/2020/059 and HIM/2022/040).

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