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Biocompatibilities and biodegradation of poly(3-hydroxybutyrate-co-3-hydroxyvalerate)s produced by a model metabolic reaction-based system.

Napathorn SC - BMC Microbiol. (2014)

Bottom Line: The diad and triad sequence distributions indicated that the PHBVs produced via the fed-batch cultivation using two different feed systems resulted in two types of copolymers: random PHBVs and putative block PHBVs.The production of IL-8, which is induced by PHB and PHBVs, may be used to improve and enhance the wound-healing process because of deficiencies of IL-8 in the wound area, particularly in problematic wounds.Taken together, the results support the use of PHB and the random and putative block PHBVs produced in this study as potential biomaterials in tissue engineering applications for connective tissue, bone and dermal fibroblast reconstruction.

View Article: PubMed Central - PubMed

Affiliation: Department of Microbiology, Faculty of Science, Chulalongkorn University, Phayathai Road, Patumwan, Bangkok, 10330, Thailand. suchada.cha@chula.ac.th.

ABSTRACT

Background: This study evaluated the biocompatibilities of random and putative block poly(3-hydroxybutyrate-co-3-hydroxyvalerate)s (PHBVs) produced by a metabolic reaction-based system. The produced PHBVs were fractionated, and the copolymer sequence distributions were analyzed using (1)H and (13)C NMR spectroscopy. The thermal properties were analyzed using differential scanning calorimetry (DSC). Mechanical tests were conducted using a universal testing machine. The in vitro cytotoxicities of films composed of random PHBVs and putative block PHBVs were investigated against three types of mammalian cells. The surfaces of the copolymer films and the morphologies of the cells were qualitatively monitored using scanning electron microscopy (SEM) and atomic force microscopy (AFM).

Results: Films composed of poly(3-hydroxybutyrate) (PHB), random PHBVs, putative block PHBVs, polystyrene and polyvinylchloride were prepared and characterized. The diad and triad sequence distributions indicated that the PHBVs produced via the fed-batch cultivation using two different feed systems resulted in two types of copolymers: random PHBVs and putative block PHBVs. The monomer compositions and sequence distributions strongly affected the thermal and mechanical properties. The mechanical integrity and characteristics of the film surfaces changed with the HV content. Notably, the random PHBVs possessed different mechanical properties than the putative block PHBVs. The biocompatibilities of these films were evaluated in vitro against three types of mammalian cells: L292 mouse connective tissue, human dermal fibroblast and Saos-2 human osteosarcoma cells. None of the PHBV films exhibited cytotoxic responses to the three types of mammalian cells. Erosion of the PHA film surfaces was observed by scanning electron microscopy and atomic force microscopy. The production of transforming growth factor-β-1 and interleukin-8 was also examined with regards to the usefulness of PHB and PHBV as biomaterials for regenerative tissue. The production of IL-8, which is induced by PHB and PHBVs, may be used to improve and enhance the wound-healing process because of deficiencies of IL-8 in the wound area, particularly in problematic wounds.

Conclusion: Taken together, the results support the use of PHB and the random and putative block PHBVs produced in this study as potential biomaterials in tissue engineering applications for connective tissue, bone and dermal fibroblast reconstruction.

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The AFM images of human dermal fibroblasts cultured on a film sheet composed of random PHBV with 23% 3HV content; (A) 2-D image and (B) 3-D image of the original surface of the film sheet before cell cultivation for one month; (C) 2-D image and (D) 3-D image of the same film sheet after cell cultivation for one month.
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Fig7: The AFM images of human dermal fibroblasts cultured on a film sheet composed of random PHBV with 23% 3HV content; (A) 2-D image and (B) 3-D image of the original surface of the film sheet before cell cultivation for one month; (C) 2-D image and (D) 3-D image of the same film sheet after cell cultivation for one month.

Mentions: To evaluate these changes, tapping mode AFM measurements were conducted using a scanning probe microscope(SPM), the NanoScope. The results are shown in Figures 7 and 8. Figure 7A and B show the original surface of random PHBVs with 23% 3HV. The film possesses a smooth surface. However, after culturing human dermal fibroblast cells on the surfaces of the film sheets for one month, as shown in Figure 7C and D, similar erosions of the film surface were observed as that in the SEM analysis. Figure 8A and B show the original surface of block PHBV with 19% 3HV. The film surface of the block PHBV was rich with crystalline regions and rougher than the random PHBV surface with a similar 3HV content. Film surface erosion was not observed even after culturing human dermal fibroblast cells on the surface of film sheets for one month, as shown in Figure 8C and D. The cells attached on the surface are shown in white. Similar observations were made for L292 mouse connective tissue and Saos-2 human osteosarcoma cells.Figure 7


Biocompatibilities and biodegradation of poly(3-hydroxybutyrate-co-3-hydroxyvalerate)s produced by a model metabolic reaction-based system.

Napathorn SC - BMC Microbiol. (2014)

The AFM images of human dermal fibroblasts cultured on a film sheet composed of random PHBV with 23% 3HV content; (A) 2-D image and (B) 3-D image of the original surface of the film sheet before cell cultivation for one month; (C) 2-D image and (D) 3-D image of the same film sheet after cell cultivation for one month.
© Copyright Policy - open-access
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4279891&req=5

Fig7: The AFM images of human dermal fibroblasts cultured on a film sheet composed of random PHBV with 23% 3HV content; (A) 2-D image and (B) 3-D image of the original surface of the film sheet before cell cultivation for one month; (C) 2-D image and (D) 3-D image of the same film sheet after cell cultivation for one month.
Mentions: To evaluate these changes, tapping mode AFM measurements were conducted using a scanning probe microscope(SPM), the NanoScope. The results are shown in Figures 7 and 8. Figure 7A and B show the original surface of random PHBVs with 23% 3HV. The film possesses a smooth surface. However, after culturing human dermal fibroblast cells on the surfaces of the film sheets for one month, as shown in Figure 7C and D, similar erosions of the film surface were observed as that in the SEM analysis. Figure 8A and B show the original surface of block PHBV with 19% 3HV. The film surface of the block PHBV was rich with crystalline regions and rougher than the random PHBV surface with a similar 3HV content. Film surface erosion was not observed even after culturing human dermal fibroblast cells on the surface of film sheets for one month, as shown in Figure 8C and D. The cells attached on the surface are shown in white. Similar observations were made for L292 mouse connective tissue and Saos-2 human osteosarcoma cells.Figure 7

Bottom Line: The diad and triad sequence distributions indicated that the PHBVs produced via the fed-batch cultivation using two different feed systems resulted in two types of copolymers: random PHBVs and putative block PHBVs.The production of IL-8, which is induced by PHB and PHBVs, may be used to improve and enhance the wound-healing process because of deficiencies of IL-8 in the wound area, particularly in problematic wounds.Taken together, the results support the use of PHB and the random and putative block PHBVs produced in this study as potential biomaterials in tissue engineering applications for connective tissue, bone and dermal fibroblast reconstruction.

View Article: PubMed Central - PubMed

Affiliation: Department of Microbiology, Faculty of Science, Chulalongkorn University, Phayathai Road, Patumwan, Bangkok, 10330, Thailand. suchada.cha@chula.ac.th.

ABSTRACT

Background: This study evaluated the biocompatibilities of random and putative block poly(3-hydroxybutyrate-co-3-hydroxyvalerate)s (PHBVs) produced by a metabolic reaction-based system. The produced PHBVs were fractionated, and the copolymer sequence distributions were analyzed using (1)H and (13)C NMR spectroscopy. The thermal properties were analyzed using differential scanning calorimetry (DSC). Mechanical tests were conducted using a universal testing machine. The in vitro cytotoxicities of films composed of random PHBVs and putative block PHBVs were investigated against three types of mammalian cells. The surfaces of the copolymer films and the morphologies of the cells were qualitatively monitored using scanning electron microscopy (SEM) and atomic force microscopy (AFM).

Results: Films composed of poly(3-hydroxybutyrate) (PHB), random PHBVs, putative block PHBVs, polystyrene and polyvinylchloride were prepared and characterized. The diad and triad sequence distributions indicated that the PHBVs produced via the fed-batch cultivation using two different feed systems resulted in two types of copolymers: random PHBVs and putative block PHBVs. The monomer compositions and sequence distributions strongly affected the thermal and mechanical properties. The mechanical integrity and characteristics of the film surfaces changed with the HV content. Notably, the random PHBVs possessed different mechanical properties than the putative block PHBVs. The biocompatibilities of these films were evaluated in vitro against three types of mammalian cells: L292 mouse connective tissue, human dermal fibroblast and Saos-2 human osteosarcoma cells. None of the PHBV films exhibited cytotoxic responses to the three types of mammalian cells. Erosion of the PHA film surfaces was observed by scanning electron microscopy and atomic force microscopy. The production of transforming growth factor-β-1 and interleukin-8 was also examined with regards to the usefulness of PHB and PHBV as biomaterials for regenerative tissue. The production of IL-8, which is induced by PHB and PHBVs, may be used to improve and enhance the wound-healing process because of deficiencies of IL-8 in the wound area, particularly in problematic wounds.

Conclusion: Taken together, the results support the use of PHB and the random and putative block PHBVs produced in this study as potential biomaterials in tissue engineering applications for connective tissue, bone and dermal fibroblast reconstruction.

Show MeSH
Related in: MedlinePlus