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. 2020 Mar 21;13(6):1433.
doi: 10.3390/ma13061433.

Bacterial Cellulose-Modified Polyhydroxyalkanoates Scaffolds Promotes Bone Formation in Critical Size Calvarial Defects in Mice

Ada Codreanu  1 , Cornel Balta  2 , Hildegard Herman  2 , Coralia Cotoraci  1 , Ciprian Valentin Mihali  1 , Nicoleta Zurbau  1 , Catalin Zaharia  3 , Maria Rapa  4 , Paul Stanescu  3 , Ionut-Cristian Radu  3 , Eugeniu Vasile  5 , George Lupu  6 , Bianca Galateanu  7 , Anca Hermenean  1   2
Affiliations

Bacterial Cellulose-Modified Polyhydroxyalkanoates Scaffolds Promotes Bone Formation in Critical Size Calvarial Defects in Mice

Ada Codreanu et al. Materials (Basel). .

Erratum in

Abstract

Bone regeneration is a claim challenge in addressing bone defects with large tissue deficits, that involves bone grafts to support the activity. In vitro biocompatibility of the bacterial cellulose-modified polyhydroxyalkanoates (PHB/BC) scaffolds and its osteogenic potential in critical-size mouse calvaria defects had been investigated. Bone promotion and mineralization were analyzed by biochemistry, histology/histomorphometry, X-ray analysis and immunofluorescence for highlighting osteogenesis markers. In summary, our results showed that PHB/BC scaffolds are able to support 3T3-L1 preadipocytes proliferation and had a positive effect on in vivo osteoblast differentiation, consequently inducing new bone formation after 20 weeks post-implantation. Thus, the newly developed PHB/BC scaffolds could turn out to be suitable biomaterials for the bone tissue engineering purpose.

Keywords: bacterial cellulose; bone tissue engineering; in vivo tests; polyhydroxyalkanoates.

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Conflict of interest statement

The authors declare no conflict of interest

Figures

Figure 1
Figure 1
Surgical procedure. (a) 5-mm critical size calvaria defect (b,c) Placement of the PHB/BC scaffold (d) Closure of the surgical site.
Figure 2
Figure 2
SEM microphotographs of PHB/BC scaffolds compared with plasticized PHB. (A) plasticized PHB; (B) (PHB:TBC)_BC1; (C) (PHB:TBC)_BC2.
Figure 3
Figure 3
FTIR spectra of neat PHB and PHB blends (a) Neat PHB; (b) PHB:TBC; (c) (PHB:TBC)_BC1; (d) (PHB/:TBC)_BC2.
Figure 4
Figure 4
Fluorescence microscopy micrographs of DAPI stained nuclei (blue) in 3T3-L1 preadipocytes cultivated for 24 h and 5 days on the scaffolds surface. Scale 100 μm.
Figure 5
Figure 5
Graphic representation of the spectrophotometric data obtained by the MTT assay revealing the 3T3-L1 preadipocytes viability and proliferation potential after 24 h and 5 days.
Figure 6
Figure 6
ALP activity in mice calvaria defects implanted with PHB/BC scaffolds after 3 days, 4 weeks and 20 weeks post-implantation. ### p < 0.0001 scaffods vs negative control (calvaria defect) at 3 days; **** p < 0.0001 3 days vs. 4 weeks (for each scaffold) si **** p < 0.0001 3 days vs. 20 weeks (for each scaffold).
Figure 7
Figure 7
Radiographs of the in vivo bone samples taken after 3 days, 4 and 20 weeks of BC/PHB scaffold implantation. Radiological analysis showing gradual increase in bone formation in BC/PHB matrix compared to PHB matrix. Magnification ×2.5.
Figure 8
Figure 8
Histology and histomorphometry of the PHB/BC scaffolds after 3 days, 4 and 20 weeks post-implantation (A) Histology of the PHB, (PHB:TBC)_BC1 and (PHB:TBC)_BC2 after 3 days, 4 and 20 weeks of PHB/BC scaffolds implantation; symbols: scaffold (Sc); granulation tissue (GT); fibroconnective tissue (Fc); capillary (arrow); new bone (NB); old bone (OB); Masson Goldner trichrome stain. Scale 50 μm. (B) Histomorphometry showing bone recovery in calvaria defects implanted with CHB/BCs scaffolds after 4 and 20 weeks post-implantation, compared to negative control (only defect).
Figure 9
Figure 9
Immunofluorescent detection of osterix (OSX) in the repaired calvaria (in red) after 3 days, 4 and 20 weeks of PHB/BC scaffolds implantation. Cell nuclei were stained with DAPI (in blue). ×63 oil-immersion objective.
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References

    1. Dimitriou R., Jones E., McGonagle D., Giannoudis P.V. Bone regeneration: Current concepts and future directions. BMC Med. 2011;9:66. doi: 10.1186/1741-7015-9-66. - DOI - PMC - PubMed
    1. Roberts T.T., Rosenbaum A.J. Bone grafts, bone substitutes and orthobiologics. Organogenesis. 2012;8:114–124. doi: 10.4161/org.23306. - DOI - PMC - PubMed
    1. Dhandayuthapani B., Yoshida Y., Maekawa T., Kumar D.S. Polymeric Scaffolds in Tissue Engineering Application: A Review. [(accessed on 3 February 2020)]. Available online: https://www.hindawi.com/journals/ijps/2011/290602/
    1. Oliveira Barud H.G., Barud H.d.S., Cavicchioli M., do Amaral T.S., de Oliveira Junior O.B., Santos D.M., Petersen A.L.d.O.A., Celes F., Borges V.M., de Oliveira C.I., et al. Preparation and characterization of a bacterial cellulose/silk fibroin sponge scaffold for tissue regeneration. Carbohydr. Polym. 2015;128:41–51. doi: 10.1016/j.carbpol.2015.04.007. - DOI - PubMed
    1. de Oliveira Barud H.G., da Silva R.R., da Silva Barud H., Tercjak A., Gutierrez J., Lustri W.R., de Oliveira O.B., Ribeiro S.J.L. A multipurpose natural and renewable polymer in medical applications: Bacterial cellulose. Carbohydr. Polym. 2016;153:406–420. doi: 10.1016/j.carbpol.2016.07.059. - DOI - PubMed

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