Biocompatibility of Artificial Cornea Based on Genipin-Cross-Linked Amniotic Membrane

2021 ◽  
Author(s):  
Liu Yang ◽  
Yunchuan Li ◽  
Yanfeng Yin ◽  
Qiufen Tu ◽  
Zheng Guan ◽  
...  
Keyword(s):  
Tissue Engineering ◽  
Inflammatory Response ◽  
Amniotic Membrane ◽  
Artificial Organs ◽  
Epithelial Tissue ◽  
Human Umbilical Cord ◽  
Artificial Cornea ◽  
Corneal Epithelial ◽  
Engineering Technique

Tissue engineering is a method of constructing seeding cells and artificial materials as the cytoskeleton in vitro, in order to fabricating artificial organs and tissues. Reconstruction of corneal epithelial tissue in vitro by tissue engineering technique brought hope to the corneal blind patients. In this study, we used human umbilical cord mesenchymal stem cells (hUC-MSCs) as seeding cells, and cross-linked amniotic membrane by genipin as the cytoskeleton to reconstructing corneal epithelial tissue in vitro. In addition, we tested the tenacity, hardness, degradation speed, cytocompatibility and inflammatory response in preclinical application of this new artificial material, for the purpose of finding a new approach of modifying amniotic membrane close to the feature of natural cornea. As a result, the best cross-link condition-1.0% genipin cross-linked with amniotic membrane under 45°C for 24 hr could improve the physical character of natural amniotic membrane. Genipin cross-linking makes amniotic membrane and seeded hUC-MSCs has better cytocompatibility and lower inflammatory response in preclinical application.

scholarly journals Decellularized Human Stromal Lenticules Combine with Corneal Epithelial-Like Cells: A New Resource for Corneal Tissue Engineering

2019 ◽  
Vol 2019 ◽  
pp. 1-10 ◽  
Author(s):  
Shuai Qin ◽  
Shuai Zheng ◽  
Bing Qi ◽  
Rui Guo ◽  
Guanghui Hou
Keyword(s):  
Tissue Engineering ◽  
Induction Medium ◽  
Nuclear Materials ◽  
Corneal Tissue ◽  
Corneal Epithelial ◽  
Human Ipscs ◽  
Immunological Rejection ◽  
Lenticule Extraction ◽  
Induced Pluripotent

The lack of donor corneal tissue or the immunological rejection remains a challenge for individuals with limbal stem cell deficiency (LSCD) who are treated with keratoplasty. Numerous lenticules which were extracted by small incision lenticule extraction (SMILE) appear to be useful materials for keratoplasty. In order to reduce the incidence of allograft rejection, lenticules would be decellularized. Lenticules which were treated with liquid nitrogen and nucleases had no cellular and nuclear materials remained. Human induced pluripotent stem cells (iPSCs) can be generated from the patient who requires keratoplasty, offering an autologous alternative and eliminating the risk of graft rejection. We found that BMP-4, RA, N-2 supplement, hEGF, B27, decellularized human stromal lenticules, conditioned medium, or induction medium promoted the differentiation of human iPSCs with high purity. The results showed that human iPSCs cultured for 4 days in differentiation medium A, 14 days in condition medium, and 1 week in induction medium on decellularized human stromal lenticules developed markedly higher expression of the markers P63, CK3, and CK12 than did those in the other methods. The level of gene expression of the epithelial and pluripotency markers and analysis by scanning electron microscopy and immunohistochemistry also showed successful differentiation. After inducing differentiation in vitro, corneal epithelial-like cells were induced. In the study, we investigated the possibility of a new resource for corneal tissue engineering.

scholarly journals Anisotropic cytocompatible electrospun scaffold for tendon tissue engineering elicits limited inflammatory response in vitro

2018 ◽  
Vol 33 (1) ◽  
pp. 127-139 ◽  
Author(s):  
Andrea Fotticchia ◽  
David Musson ◽  
Cristina Lenardi ◽  
Emrah Demirci ◽  
Yang Liu
Keyword(s):  
Tissue Engineering ◽  
Inflammatory Response ◽  
Social Impact ◽  
Polymer Processing ◽  
High Recurrence Rate ◽  
Tendon Tissue ◽  
Tendon Tissue Engineering ◽  
Tendon Tears

Tendon tears are a relevant concern for today’s national health systems because of their social impact and high recurrence rate. The current gold standard for fixing tendon tears is surgical repair; however, this strategy is not able to fully re-establish tendon integrity and functionality. Tissue engineering approaches aim at promoting tissue regeneration by delivering the opportune signals to the injured site combining biomaterials, cells and biochemical cues. Electrospinning is currently one of the most versatile polymer processing techniques that allows manufacturing of nano- and micro-fibres substrates. Such fibrous morphology is deemed to be an ideal substrate to convey topographical cues to cells. Here we evaluated the potential of polycaprolactone processed by means of electrospinning technology for tendon tissue engineering. Fibrous free-of-defects substrate with random and aligned fibres were successfully fabricated. Rat tenocytes were used to assess the cytocompatibility of the substrates for application as tendon tissue engineered devices. Tenocytes were able to proliferate and adapt to the substrates topography acquiring an elongated morphology, which is the precondition for oriented collagen deposition, when seeded on aligned fibres. Real time Polymerase Chain Reaction (Rt-PCR) also revealed the overall maintenance of tenocyte phenotype over 7 days culture. To verify suitability for in vivo implantation, the level of inflammatory cytokine genes expressed by THP-1 cells cultured in presence of electrospun polycaprolactone substrates was evaluated. Inflammatory response was limited. The novel preliminary in vitro work presented herein showing tenocytes compatibility and limited inflammatory cytokines synthesis suggests that electrospun polycaprolactone may be taken into consideration as substrate for tendon healing applications.

scholarly journals Layer-by-layer decorated herbal cell compatible scaffolds for bone tissue engineering: A synergistic effect of graphene oxide and Cissus quadrangularis

2020 ◽  
Vol 35 (1) ◽  
pp. 57-73 ◽  
Author(s):  
Shivaji Kashte ◽  
RK Sharma ◽  
Sachin Kadam
Keyword(s):  
Tissue Engineering ◽  
Graphene Oxide ◽  
Synergistic Effect ◽  
Bone Tissue Engineering ◽  
Bone Tissue ◽  
Cellular Proliferation ◽  
Human Umbilical Cord ◽  
Layer By Layer ◽  
Cissus Quadrangularis

Among various bone regenerative and repair methods, use of osteoinductive scaffold as bone grafts/substitute has gained wide importance worldwide. To develop such osteoinductive scaffold that is more natural and which spontaneously stimulates osteoblast formation without any differentiation media, we prepared electrospun poly ε-caprolactone scaffold which is further modified by means of layer-by-layer method using Cissus quadrangularis callus culture extract and graphene oxide (PCL-GO-CQ). The modified PCL-GO-CQ scaffold was compared with plain poly ε-caprolactone scaffold and poly ε-caprolactone coated only with graphene oxide. Physical properties, such as roughness, wettability, yield strength and tensile strength, of PCL-GO-CQ scaffold were found to be superior. Also, PCL-GO-CQ scaffold showed more in vitro cell compatibility with enhanced cellular proliferation on its surface. Presence of graphene oxide and Cissus quadrangularis callus in scaffold helped in the differentiation of human umbilical cord Wharton’s jelly-derived mesenchymal stem cells into osteogenic lineage without any differentiation media in less than 20 days. The synergistic effect of Cissus quadrangularis callus extract and graphene oxide in PCL-GO-CQ scaffold enhanced osteoblastic differentiation, osteoconduction and osteoinduction potential of scaffolds making them highly potential in bone regeneration and bone tissue engineering applications.

scholarly journals Umbilical Cord Mesenchymal Stem Cells-derived Exosomes Deliver Mir-21 to Accelerate Corneal Epithelial Wound Healing Through PTEN/PI3K/Akt pathway

2021 ◽  
Author(s):  
Xuran Li ◽  
Xiaolong Liu ◽  
Yanyan Zhang ◽  
Zhiyu Liu ◽  
Xinyue Li ◽  
...  
Keyword(s):  
Stem Cells ◽  
Wound Healing ◽  
Mesenchymal Stem Cells ◽  
Umbilical Cord ◽  
Wound Repair ◽  
Recipient Cell ◽  
Human Umbilical Cord ◽  
Corneal Epithelial ◽  
Epithelial Wound Healing

Abstract BackgroundRapid restoration of corneal epithelium integrity after injury is particularly important for preserving corneal transparency and vision. Mesenchymal stem cells (MSCs) can be taken into account as the promising regenerative therapeutics for improvement of wound healing processes based on the variety of the effective components. The extracellular vesicles form MSCs, especially exosomes, has been considered as important paracrine mediators though transferring microRNAs into recipient cell. This study investigated the mechanism of human umbilical cord MSC-derived exosomes (HUMSC-exosomes) on corneal epithelial wound healing.MethodsExosomes extracted from HUMSCs were identified by transmission electron microscopy, nanoparticle tracking analysis, and Western blot. Corneal fluorescein staining and histological staining were evaluated in a corneal mechanical wound model. Changes in HCECs proliferation after HUMSC-exosomes or miR-21 mimic treatment were evaluated by CCK-8 and EdU assays, while migration was assessed by in vitro scratch wound assay. Full-length transcriptome sequencing was performed to identify the differentially expressed genes associated with HUMSC-exosomes treatment, followed by validation via real-time PCR and Western blot.ResultsThe exosomes derived from HUMSCs can significantly promote corneal epithelial cells proliferation, migration in vitro and accelerate corneal epithelial wound healing in vivo. Similar effects were obtained after miR-21 transfection, while the beneficial effects of HUMSC-exosomes were partially negated by miR-21 knockdown. Results also show that the benefits are associated with decreased PTEN level and activated the PI3K/Akt signaling pathway in HCECs.ConclusionsHUMSC-exosomes could accelerate the recovery of corneal epithelial wounds though restraining PTEN by transferring miR-21, and may represent a promising novel therapeutic agent for corneal wound repair.

scholarly journals Anandamide Concentration-Dependently Modulates Toll-Like Receptor 3 Agonism or UVB-Induced Inflammatory Response of Human Corneal Epithelial Cells

2021 ◽  
Vol 22 (15) ◽  
pp. 7776
Author(s):  
Ágnes Angyal ◽  
Zsófia Pénzes ◽  
Shahrzad Alimohammadi ◽  
Dorottya Horváth ◽  
Lili Takács ◽  
...  
Keyword(s):  
Epithelial Cells ◽  
Inflammatory Response ◽  
Toll Like Receptor ◽  
Corneal Epithelial Cells ◽  
Corneal Epithelial ◽  
Uvb Irradiation ◽  
Human Corneal Epithelial Cells ◽  
Anti Inflammatory

Photodamage-induced and viral keratitis could benefit from treatment with novel nonsteroid anti-inflammatory agents. Therefore, we determined whether human corneal epithelial cells (HCECs) express members of the endocannabinoid system (ECS), and examined how the endocannabinoid anandamide (AEA, N-arachidonoyl ethanolamine) influences the Toll-like receptor 3 (TLR3) agonism- or UVB irradiation-induced inflammatory response of these cells. Other than confirming the presence of cannabinoid receptors, we show that endocannabinoid synthesizing and catabolizing enzymes are also expressed in HCECs in vitro, as well as in the epithelial layer of the human cornea in situ, proving that they are one possible source of endocannabinoids. p(I:C) and UVB irradiation was effective in promoting the transcription and secretion of inflammatory cytokines. Surprisingly, when applied alone in 100 nM and 10 μM, AEA also resulted in increased pro-inflammatory cytokine production. Importantly, AEA further increased levels of these cytokines in the UVB model, whereas its lower concentration partially prevented the transcriptional effect of p(I:C), while not decreasing the p(I:C)-induced cytokine release. HCECs express the enzymatic machinery required to produce endocannabinoids both in vitro and in situ. Moreover, our data show that, despite earlier reports about the anti-inflammatory potential of AEA in murine cornea, its effects on the immune phenotype of human corneal epithelium may be more complex and context dependent.

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