Modulation of Macrophage Phenotype, Maturation, and Graft Integration through Chondroitin Sulfate Cross-Linking to Decellularized Cornea
- Juhi ChakrabortyJuhi ChakrabortyRegenerative Engineering Laboratory, Department of Textile Technology, Indian Institute of Technology Delhi, New Delhi 110016, IndiaMore by Juhi Chakraborty
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- Subhadeep RoySubhadeep RoyRegenerative Engineering Laboratory, Department of Textile Technology, Indian Institute of Technology Delhi, New Delhi 110016, IndiaMore by Subhadeep Roy
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- Sumit MurabSumit MurabRegenerative Engineering Laboratory, Department of Textile Technology, Indian Institute of Technology Delhi, New Delhi 110016, IndiaMore by Sumit Murab
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- Raghav RavaniRaghav RavaniDr Rajendra Prasad Centre for Ophthalmic Sciences, All India Institute of Medical Sciences, New Delhi 110016, IndiaMore by Raghav Ravani
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- Kulwinder KaurKulwinder KaurRegenerative Engineering Laboratory, Department of Textile Technology, Indian Institute of Technology Delhi, New Delhi 110016, IndiaMore by Kulwinder Kaur
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- Saranya DeviSaranya DeviDr Rajendra Prasad Centre for Ophthalmic Sciences, All India Institute of Medical Sciences, New Delhi 110016, IndiaMore by Saranya Devi
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- Divya SinghDivya SinghDr Rajendra Prasad Centre for Ophthalmic Sciences, All India Institute of Medical Sciences, New Delhi 110016, IndiaMore by Divya Singh
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- Shubhangini SharmaShubhangini SharmaStem Cell Facility, All India Institute of Medical Sciences, New Delhi 110016, IndiaMore by Shubhangini Sharma
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- Sujata MohantySujata MohantyStem Cell Facility, All India Institute of Medical Sciences, New Delhi 110016, IndiaMore by Sujata Mohanty
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- Amit Kumar DindaAmit Kumar DindaDepartment of Pathology, All India Institute of Medical Sciences, New Delhi 110016, IndiaMore by Amit Kumar Dinda
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- Radhika TandonRadhika TandonDr Rajendra Prasad Centre for Ophthalmic Sciences, All India Institute of Medical Sciences, New Delhi 110016, IndiaMore by Radhika Tandon
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- Sourabh Ghosh*Sourabh Ghosh*E-mail: [email protected]Regenerative Engineering Laboratory, Department of Textile Technology, Indian Institute of Technology Delhi, New Delhi 110016, IndiaMore by Sourabh Ghosh
Abstract
Decellularized corneas obtained from other species have gained intense popularity in the field of tissue engineering due to its role to serve as an alternative to the limited availability of high-quality donor tissues. However, the decellularized cornea is found to evoke an immune response inspite of the removal of the cellular contents and antigens due to the distortion of the collagen fibrils that exposes certain antigenic sites, which often lead to graft rejection. Therefore, in this study we tested the hypothesis that cross-linking the decellularized corneas with chondroitin sulfate may help in restoring the distorted conformationation changes of fibrous matrix and thus help in reducing the occurrence of graft rejection. Cross-linking of the decellularized cornea with oxidized chondroitin sulfate was validated by ATR-FTIR analysis. An in vitro immune response study involving healthy monocytes and differentiated macrophages with their surface marker analysis by pHrodo red, Lysotracker red, ER tracker, and CD63, LAMP-2 antibodies confirmed that the cross-linked decellularized matrices elicited the least immune response compared to the decellularized ones. We implanted three sets of corneal scaffolds obtained from goat, i.e., native, decellularized, and decellularized corneas conjugated with chondroitin sulfate into the rabbit stroma. Histology analysis, three months after implantation into the rabbit corneal stromal region, confirmed the restoration of the collagen fibril conformation and the migration of cells to the implanted constructs, affirming proper graft integration. Hence we conclude that the chondroitin sulfate cross-linked decellularized corneal matrix may serve as an efficient alternative to the allograft and human cadaveric corneas.
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