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Contribution of fibroblasts to the mechanical stability of in vitro engineered dermal-like tissue through extracellular matrix deposition.

Nair RP, Joseph J, Harikrishnan VS, Krishnan VK, Krishnan L - Biores Open Access (2014)

Bottom Line: To enhance cell growth, synthetic PLGC was incorporated with a fibrin-based biomimetic composite.The loss of mechanical strength consequent to in vitro polymer degradation seemed to be balanced owing to the ECM deposition.Similar patterns of in vivo and in vitro degradation were observed during subcutaneous implantation and fibroblast culture, respectively.

View Article: PubMed Central - PubMed

Affiliation: Thrombosis Research Unit, Biomedical Technology Wing, Sree Chitra Tirunal Institute for Medical Sciences and Technology , Trivandrum, India .

ABSTRACT
Tissue-engineered skin with mechanical and biological properties that match the native tissue could be a valuable graft to treat non-healing chronic wounds. Fibroblasts grown on a suitable biodegradable scaffold are a feasible strategy for the development of a dermal substitute above which epithelialization may occur naturally. Cell growth and phenotype maintenance are crucial to ensure the functional status of engineered tissue. In this study, an electrospun biodegradable polymer scaffold composed of a terpolymer PLGC [poly(lactide-glycolide-caprolactone)] with appropriate mechanical strength was used as a scaffold so that undesirable contraction of the wound could be prevented when it was implanted. To enhance cell growth, synthetic PLGC was incorporated with a fibrin-based biomimetic composite. The efficacy of the hybrid scaffold was evaluated by comparing it with bare PLGC in terms of fibroblast growth potential, extracellular matrix (ECM) deposition, polymer degradation, and mechanical strength. A significant increase was observed in fibroblast attachment, proliferation, and deposition of ECM proteins such as collagen and elastin in the hybrid scaffold. After growing fibroblasts for 20 d and 40 d, immunochemical staining of the decellularized scaffolds showed deposition of insoluble collagen and elastin on the hybrid scaffold but not on the bare scaffold. The loss of mechanical strength consequent to in vitro polymer degradation seemed to be balanced owing to the ECM deposition. Thus, tensile strength and elongation were better when cells were grown on the hybrid scaffold rather than the bare samples immersed in culture medium. Similar patterns of in vivo and in vitro degradation were observed during subcutaneous implantation and fibroblast culture, respectively. We therefore postulate that a hybrid scaffold comprising PLGC and fibrin is a potential candidate for the engineering of dermal tissue to be used in the regeneration of chronic wounds.

No MeSH data available.


Related in: MedlinePlus

Data on in vitro degradation of scaffold. (A) Change in molecular weight (weight average [Mw] and number average [Mn]) of polymer suspended in culture medium with or without fibroblast after different periods. (B) Tensile strength and percent elongation of hybrid scaffolds suspended in culture medium with or without fibroblast after different periods. Elongation was higher for cell-grown samples but was not statistically significant. Mechanical testing of cell-free scaffolds was not possible after 40 d of immersion in the medium because they were found to have lost the required dimension due to shrinkage. Data presented is average±SD (n=5). *p<0.05. The periods of analysis are marked in the x-axis.
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f5: Data on in vitro degradation of scaffold. (A) Change in molecular weight (weight average [Mw] and number average [Mn]) of polymer suspended in culture medium with or without fibroblast after different periods. (B) Tensile strength and percent elongation of hybrid scaffolds suspended in culture medium with or without fibroblast after different periods. Elongation was higher for cell-grown samples but was not statistically significant. Mechanical testing of cell-free scaffolds was not possible after 40 d of immersion in the medium because they were found to have lost the required dimension due to shrinkage. Data presented is average±SD (n=5). *p<0.05. The periods of analysis are marked in the x-axis.

Mentions: The scaffold patches without cells shrunk over time when immersed in the medium, while the cell-grown scaffolds maintained their shape and dimension. Polymer degradation was evident from the reduction in both Mw and Mn by 20 d and 40 d, more significantly in the cell grown scaffolds (Fig. 5A, B). However, the tensile strength of the cell-grown scaffolds was significantly high as compared to that of cell-free ones on 20 d (Fig. 5C); by 40 d, in spite of progression in polymer breakdown which resulted in the reduction in Mn and Mw, the tensile strength was steady (Fig. 5B vs. Fig. 5D).


Contribution of fibroblasts to the mechanical stability of in vitro engineered dermal-like tissue through extracellular matrix deposition.

Nair RP, Joseph J, Harikrishnan VS, Krishnan VK, Krishnan L - Biores Open Access (2014)

Data on in vitro degradation of scaffold. (A) Change in molecular weight (weight average [Mw] and number average [Mn]) of polymer suspended in culture medium with or without fibroblast after different periods. (B) Tensile strength and percent elongation of hybrid scaffolds suspended in culture medium with or without fibroblast after different periods. Elongation was higher for cell-grown samples but was not statistically significant. Mechanical testing of cell-free scaffolds was not possible after 40 d of immersion in the medium because they were found to have lost the required dimension due to shrinkage. Data presented is average±SD (n=5). *p<0.05. The periods of analysis are marked in the x-axis.
© Copyright Policy
Related In: Results  -  Collection

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getmorefigures.php?uid=PMC4215331&req=5

f5: Data on in vitro degradation of scaffold. (A) Change in molecular weight (weight average [Mw] and number average [Mn]) of polymer suspended in culture medium with or without fibroblast after different periods. (B) Tensile strength and percent elongation of hybrid scaffolds suspended in culture medium with or without fibroblast after different periods. Elongation was higher for cell-grown samples but was not statistically significant. Mechanical testing of cell-free scaffolds was not possible after 40 d of immersion in the medium because they were found to have lost the required dimension due to shrinkage. Data presented is average±SD (n=5). *p<0.05. The periods of analysis are marked in the x-axis.
Mentions: The scaffold patches without cells shrunk over time when immersed in the medium, while the cell-grown scaffolds maintained their shape and dimension. Polymer degradation was evident from the reduction in both Mw and Mn by 20 d and 40 d, more significantly in the cell grown scaffolds (Fig. 5A, B). However, the tensile strength of the cell-grown scaffolds was significantly high as compared to that of cell-free ones on 20 d (Fig. 5C); by 40 d, in spite of progression in polymer breakdown which resulted in the reduction in Mn and Mw, the tensile strength was steady (Fig. 5B vs. Fig. 5D).

Bottom Line: To enhance cell growth, synthetic PLGC was incorporated with a fibrin-based biomimetic composite.The loss of mechanical strength consequent to in vitro polymer degradation seemed to be balanced owing to the ECM deposition.Similar patterns of in vivo and in vitro degradation were observed during subcutaneous implantation and fibroblast culture, respectively.

View Article: PubMed Central - PubMed

Affiliation: Thrombosis Research Unit, Biomedical Technology Wing, Sree Chitra Tirunal Institute for Medical Sciences and Technology , Trivandrum, India .

ABSTRACT
Tissue-engineered skin with mechanical and biological properties that match the native tissue could be a valuable graft to treat non-healing chronic wounds. Fibroblasts grown on a suitable biodegradable scaffold are a feasible strategy for the development of a dermal substitute above which epithelialization may occur naturally. Cell growth and phenotype maintenance are crucial to ensure the functional status of engineered tissue. In this study, an electrospun biodegradable polymer scaffold composed of a terpolymer PLGC [poly(lactide-glycolide-caprolactone)] with appropriate mechanical strength was used as a scaffold so that undesirable contraction of the wound could be prevented when it was implanted. To enhance cell growth, synthetic PLGC was incorporated with a fibrin-based biomimetic composite. The efficacy of the hybrid scaffold was evaluated by comparing it with bare PLGC in terms of fibroblast growth potential, extracellular matrix (ECM) deposition, polymer degradation, and mechanical strength. A significant increase was observed in fibroblast attachment, proliferation, and deposition of ECM proteins such as collagen and elastin in the hybrid scaffold. After growing fibroblasts for 20 d and 40 d, immunochemical staining of the decellularized scaffolds showed deposition of insoluble collagen and elastin on the hybrid scaffold but not on the bare scaffold. The loss of mechanical strength consequent to in vitro polymer degradation seemed to be balanced owing to the ECM deposition. Thus, tensile strength and elongation were better when cells were grown on the hybrid scaffold rather than the bare samples immersed in culture medium. Similar patterns of in vivo and in vitro degradation were observed during subcutaneous implantation and fibroblast culture, respectively. We therefore postulate that a hybrid scaffold comprising PLGC and fibrin is a potential candidate for the engineering of dermal tissue to be used in the regeneration of chronic wounds.

No MeSH data available.


Related in: MedlinePlus