Limits...
Xenogeneic acellular conjunctiva matrix as a scaffold of tissue-engineered corneal epithelium.

Zhao H, Qu M, Wang Y, Wang Z, Shi W - PLoS ONE (2014)

Bottom Line: In the present study, we investigated the preparation of xenogeneic acellular conjunctiva matrix (aCM) and evaluated its efficacy and safety as a scaffold of tissue-engineered corneal epithelium.The corneal epithelial cells seeded on aCM formed a multilayered epithelial structure and endured longer than did those on dAM.These findings support the application of xenogeneic acellular conjunctiva matrix as a scaffold for reconstructing the ocular surface.

View Article: PubMed Central - PubMed

Affiliation: Ophthalmology Department, Yijishan Hospital of Wannan Medical College, Wuhu, Anhui, China; State Key Laboratory Cultivation Base, Shandong Provincial Key Laboratory of Ophthalmology, Shandong Eye Institute, Shandong Academy of Medical Sciences, Qingdao, Shandong, China.

ABSTRACT
Amniotic membrane-based tissue-engineered corneal epithelium has been widely used in the reconstruction of the ocular surface. However, it often degrades too early to ensure the success of the transplanted corneal epithelium when treating patients with severe ocular surface disorders. In the present study, we investigated the preparation of xenogeneic acellular conjunctiva matrix (aCM) and evaluated its efficacy and safety as a scaffold of tissue-engineered corneal epithelium. Native porcine conjunctiva was decellularized with 0.1% sodium dodecyl sulfate (SDS) for 12 h at 37°C and sterilized via γ-irradiation. Compared with native conjunctiva, more than 92% of the DNA was removed, and more than 90% of the extracellular matrix components (glycosaminoglycan and collagen) remained after the decellularization treatment. Compared with denuded amniotic membrane (dAM), the aCM possessed favorable optical transmittance, tensile strength, stability and biocompatibility as well as stronger resistance to degradation both in vitro and in vivo. The corneal epithelial cells seeded on aCM formed a multilayered epithelial structure and endured longer than did those on dAM. The aCM-based tissue-engineered corneal epithelium was more effective in the reconstruction of the ocular surface in rabbits with limbal stem cell deficiency. These findings support the application of xenogeneic acellular conjunctiva matrix as a scaffold for reconstructing the ocular surface.

Show MeSH

Related in: MedlinePlus

Macroscopic view of the acellular conjunctiva matrix.The aCM (B, E) were more transparent than the normal conjunctiva matrix (A, D). The transparent characteristics were not affected after the sterilization with γ-irradiation (C, F).
© Copyright Policy
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC4222936&req=5

pone-0111846-g001: Macroscopic view of the acellular conjunctiva matrix.The aCM (B, E) were more transparent than the normal conjunctiva matrix (A, D). The transparent characteristics were not affected after the sterilization with γ-irradiation (C, F).

Mentions: The decellularized and sterilized conjunctiva matrix maintained its transparent characteristics during the prepared period (Fig. 1). Compared with the native conjunctiva, no intact cells were observed in the aCM with H.E. or Hochest staining (Fig. 2). Ultrastructural observation showed that the collagen fibrils of the aCM were tight and regular, with no cellular debris in the aCM (Fig. 2). The collagen fibrils of native conjunctiva were aligned slightly more tightly than were those of the aCM (Fig. 2). The α-gal detection showed that there was no α-gal antigen in the aCM compared with the large amount of α-gal staining in the native conjunctiva, which was visualized as brown particles (Fig. 2). The levels of DNA in the aCM and native conjunctiva were 1.01±0.12 µg/mg and 12.46±1.51 µg/mg dry weight, respectively, indicating that the DNA content of aCM was much lower than that of native conjunctiva (n = 5, p<0.05). The GAG levels in aCM and native conjunctiva were 48.63±3.40 µg/mg and 52.09±4.46 µg/mg, respectively; the hydroxyproline levels were 74.87±7.09 µg/mg and 82.07±4.95 µg/mg, respectively (n = 5, p>0.05).


Xenogeneic acellular conjunctiva matrix as a scaffold of tissue-engineered corneal epithelium.

Zhao H, Qu M, Wang Y, Wang Z, Shi W - PLoS ONE (2014)

Macroscopic view of the acellular conjunctiva matrix.The aCM (B, E) were more transparent than the normal conjunctiva matrix (A, D). The transparent characteristics were not affected after the sterilization with γ-irradiation (C, F).
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4222936&req=5

pone-0111846-g001: Macroscopic view of the acellular conjunctiva matrix.The aCM (B, E) were more transparent than the normal conjunctiva matrix (A, D). The transparent characteristics were not affected after the sterilization with γ-irradiation (C, F).
Mentions: The decellularized and sterilized conjunctiva matrix maintained its transparent characteristics during the prepared period (Fig. 1). Compared with the native conjunctiva, no intact cells were observed in the aCM with H.E. or Hochest staining (Fig. 2). Ultrastructural observation showed that the collagen fibrils of the aCM were tight and regular, with no cellular debris in the aCM (Fig. 2). The collagen fibrils of native conjunctiva were aligned slightly more tightly than were those of the aCM (Fig. 2). The α-gal detection showed that there was no α-gal antigen in the aCM compared with the large amount of α-gal staining in the native conjunctiva, which was visualized as brown particles (Fig. 2). The levels of DNA in the aCM and native conjunctiva were 1.01±0.12 µg/mg and 12.46±1.51 µg/mg dry weight, respectively, indicating that the DNA content of aCM was much lower than that of native conjunctiva (n = 5, p<0.05). The GAG levels in aCM and native conjunctiva were 48.63±3.40 µg/mg and 52.09±4.46 µg/mg, respectively; the hydroxyproline levels were 74.87±7.09 µg/mg and 82.07±4.95 µg/mg, respectively (n = 5, p>0.05).

Bottom Line: In the present study, we investigated the preparation of xenogeneic acellular conjunctiva matrix (aCM) and evaluated its efficacy and safety as a scaffold of tissue-engineered corneal epithelium.The corneal epithelial cells seeded on aCM formed a multilayered epithelial structure and endured longer than did those on dAM.These findings support the application of xenogeneic acellular conjunctiva matrix as a scaffold for reconstructing the ocular surface.

View Article: PubMed Central - PubMed

Affiliation: Ophthalmology Department, Yijishan Hospital of Wannan Medical College, Wuhu, Anhui, China; State Key Laboratory Cultivation Base, Shandong Provincial Key Laboratory of Ophthalmology, Shandong Eye Institute, Shandong Academy of Medical Sciences, Qingdao, Shandong, China.

ABSTRACT
Amniotic membrane-based tissue-engineered corneal epithelium has been widely used in the reconstruction of the ocular surface. However, it often degrades too early to ensure the success of the transplanted corneal epithelium when treating patients with severe ocular surface disorders. In the present study, we investigated the preparation of xenogeneic acellular conjunctiva matrix (aCM) and evaluated its efficacy and safety as a scaffold of tissue-engineered corneal epithelium. Native porcine conjunctiva was decellularized with 0.1% sodium dodecyl sulfate (SDS) for 12 h at 37°C and sterilized via γ-irradiation. Compared with native conjunctiva, more than 92% of the DNA was removed, and more than 90% of the extracellular matrix components (glycosaminoglycan and collagen) remained after the decellularization treatment. Compared with denuded amniotic membrane (dAM), the aCM possessed favorable optical transmittance, tensile strength, stability and biocompatibility as well as stronger resistance to degradation both in vitro and in vivo. The corneal epithelial cells seeded on aCM formed a multilayered epithelial structure and endured longer than did those on dAM. The aCM-based tissue-engineered corneal epithelium was more effective in the reconstruction of the ocular surface in rabbits with limbal stem cell deficiency. These findings support the application of xenogeneic acellular conjunctiva matrix as a scaffold for reconstructing the ocular surface.

Show MeSH
Related in: MedlinePlus