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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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Related in: MedlinePlus

The production of TGF-β-1 and IL-8 from (A) L292 mouse connective tissue, (B) human dermal fibroblast and (C) Saos-2 human osteosarcoma cells when grown separately on films composed of PHB and random and putative block PHBVs for 7 days. White bars, TGF-β-1; grey bars, IL-8.
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Fig4: The production of TGF-β-1 and IL-8 from (A) L292 mouse connective tissue, (B) human dermal fibroblast and (C) Saos-2 human osteosarcoma cells when grown separately on films composed of PHB and random and putative block PHBVs for 7 days. White bars, TGF-β-1; grey bars, IL-8.

Mentions: TGF-β-1 is a growth factor that is produced by various types of cells. The amount of TGF-β-1 produced by cells is very low under normal conditions. Many studies have reported the involvement of TGF-β-1 and IL-8 during wound healing, inflammation, remodeling and bone formation [32]. In this study, L929 mouse connective tissue cells, human dermal fibroblast cells and Saos-2 human osteosarcoma cells were cultured on the surfaces of the PHB and random PHBV and putative block PHBV film sheets. The amounts of TGF-β-1 and IL-8 secreted into the cultured supernatant were determined using ELISA test kits. The control experiments were performed in the same way, except PHB or PHBV film sheets were not placed at the bottom of each well. As shown in Figure 4, 102–195, 1622–1875 and 8483–9118 pg/mLTGF-β-1 were produced from L292 mouse connective tissue (Figure 4A, white bars), human dermal fibroblasts (Figure 4B, white bars) and Saos-2 human osteosarcoma cells (Figure 4C, white bars), respectively, when grown on PHA film sheets for 7 days. However, there appears to be an obvious decrease in TGF-β-1 levels compared with their respective control levels, as shown in Figures 4A,B and C. Interestingly, opposite trends were observed when comparing the production of TGF-β-1 and IL-8 from human dermal fibroblasts (Figure 4B) and from Saos-2 human osteosarcoma cells (Figure 4C). Saos-2 osteosarcoma cells grown on PHA film sheets produced more TGF-β-1 than IL-8, whereas human dermal fibroblasts grown on PHA film sheets produced more IL-8 than TGF-β-1. Notably, as shown in Figure 4B, human dermal fibroblasts produced significantly more IL-8 than did the controls, but they produced less TGF-β-1 than did the controls. In Figure 4A, the effect of the monomeric composition in PHBV shows thatL929 mouse connective tissue cells grown on random 72%HV, random 80%HV, putative block 19%HV, putative block 41%HV and putative block 51%HV appear to have almost 2-fold higher levels of TGF-β-1 than L292 cells grown on PHB and random PHBV with low 3HVcontents. The effect of the monomeric composition in putative block PHBVs appears to be higher than that observed in random PHBVs with similar 3HV contents. However, the levels of TGF-β-1 produced from all three types of mammalian cells did not exceed the control levels.Figure 4


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 production of TGF-β-1 and IL-8 from (A) L292 mouse connective tissue, (B) human dermal fibroblast and (C) Saos-2 human osteosarcoma cells when grown separately on films composed of PHB and random and putative block PHBVs for 7 days. White bars, TGF-β-1; grey bars, IL-8.
© Copyright Policy - open-access
Related In: Results  -  Collection

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

Fig4: The production of TGF-β-1 and IL-8 from (A) L292 mouse connective tissue, (B) human dermal fibroblast and (C) Saos-2 human osteosarcoma cells when grown separately on films composed of PHB and random and putative block PHBVs for 7 days. White bars, TGF-β-1; grey bars, IL-8.
Mentions: TGF-β-1 is a growth factor that is produced by various types of cells. The amount of TGF-β-1 produced by cells is very low under normal conditions. Many studies have reported the involvement of TGF-β-1 and IL-8 during wound healing, inflammation, remodeling and bone formation [32]. In this study, L929 mouse connective tissue cells, human dermal fibroblast cells and Saos-2 human osteosarcoma cells were cultured on the surfaces of the PHB and random PHBV and putative block PHBV film sheets. The amounts of TGF-β-1 and IL-8 secreted into the cultured supernatant were determined using ELISA test kits. The control experiments were performed in the same way, except PHB or PHBV film sheets were not placed at the bottom of each well. As shown in Figure 4, 102–195, 1622–1875 and 8483–9118 pg/mLTGF-β-1 were produced from L292 mouse connective tissue (Figure 4A, white bars), human dermal fibroblasts (Figure 4B, white bars) and Saos-2 human osteosarcoma cells (Figure 4C, white bars), respectively, when grown on PHA film sheets for 7 days. However, there appears to be an obvious decrease in TGF-β-1 levels compared with their respective control levels, as shown in Figures 4A,B and C. Interestingly, opposite trends were observed when comparing the production of TGF-β-1 and IL-8 from human dermal fibroblasts (Figure 4B) and from Saos-2 human osteosarcoma cells (Figure 4C). Saos-2 osteosarcoma cells grown on PHA film sheets produced more TGF-β-1 than IL-8, whereas human dermal fibroblasts grown on PHA film sheets produced more IL-8 than TGF-β-1. Notably, as shown in Figure 4B, human dermal fibroblasts produced significantly more IL-8 than did the controls, but they produced less TGF-β-1 than did the controls. In Figure 4A, the effect of the monomeric composition in PHBV shows thatL929 mouse connective tissue cells grown on random 72%HV, random 80%HV, putative block 19%HV, putative block 41%HV and putative block 51%HV appear to have almost 2-fold higher levels of TGF-β-1 than L292 cells grown on PHB and random PHBV with low 3HVcontents. The effect of the monomeric composition in putative block PHBVs appears to be higher than that observed in random PHBVs with similar 3HV contents. However, the levels of TGF-β-1 produced from all three types of mammalian cells did not exceed the control levels.Figure 4

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