Collagen-Based Artificial Cornea for Lamellar Keratoplasty: An Early Case Report.

PURPOSE: The aim of this study was to evaluate Viscoll collagen membrane (VCM) for lamellar keratoplasty. METHODS: A 54-year-old man with grade 4 recurrent pterygium underwent lamellar keratoplasty using VCM as the graft material. Standard keratoplasty postoperative treatments, including topical antibiotic-corticosteroid and artificial tears, were administered. RESULTS: Complete graft epithelialization was achieved after 4 days of the surgery. Follow-up at 9 months showed no pterygium recurrence as well as good integration of the VCM graft into the surrounding tissues with preserved transparency. CONCLUSIONS: This case demonstrated that VCM possesses great potential as an alternative to the human donor cornea for lamellar keratoplasty; however, the application of VCM for lamellar keratoplasty still needs additional evaluation.

Matrix-Assisted Cell Transplantation for the Treatment of Limbal Stem Cell Deficiency in a Rabbit Model.

With the development of regenerative medicine in ophthalmology, the identification of cells with high proliferative potential in the limbal area has attracted the attention of ophthalmologists and offered a new option for treatment in clinical practice. Limbal stem cell deficiency (LSCD) is an identified eye disease with a difficult and negative outcome, for which the traditional treatment is keratoplasty. This study sought to evaluate the efficacy of matrix-assisted cell transplantation consisting of in vitro-cultured autologous limbal stem cells (LSCs) and type I collagen for the treatment of LSCD in rabbits. LSCD was induced in 10 rabbits by a combination of mechanical limbectomy and alkali burns. Cells were cultured on a plate for 14 days before being transferred to a collagen-based matrix for another 7 days. Rabbits were divided into two groups as follows: the experimental group (five rabbits) received matrix-assisted cell transplantation, while the control group (five rabbits) received only conservative therapy with anti-inflammatory eye drops. During the postoperative period, all rabbits were examined using slit-lamp biomicroscopy with photo-registration and fluorescent staining, impression cytology and anterior segment optical coherence tomography (AS-OCT). Rabbits were euthanized at 30 and 120 days, and their corneas were processed for histology and immunohistochemistry. As a consequence, rabbits in the experimental group demonstrated the restoration of the corneal epithelium and transparency without epithelial defects. Moreover, goblet cells were absent in the central zone of the corneal epithelium. In conclusion, our new method of treatment enhanced the corneal surface and is an effective method of treatment for LSCD in rabbits.

Corneal Stroma Regeneration with Collagen-Based Hydrogel as an Artificial Stroma Equivalent: A Comprehensive In Vivo Study.

Restoring the anatomical and functional characteristics of the cornea using various biomaterials is especially relevant in the context of a global shortage of donor tissue. Such biomaterials must be biocompatible, strong, and transparent. Here, we report a Viscoll collagen membrane with mechanical and optical properties suitable for replacing damaged stromal tissue. After removing a portion of the stroma, a Viscoll collagen membrane was implanted into the corneas of rabbits. After 6 months, the active migration of host cells into Viscoll collagen membranes was noted, with the preservation of corneal transparency in all experimental animals. Effective integration of the Viscoll collagen membrane with corneal tissue promoted nerve regeneration in vivo, as confirmed by in vivo confocal microscopy. We also demonstrated the safety and efficacy of the Viscoll collagen membrane for corneal stroma regeneration. Thus, in combination with the proposed packaging format that provides long-term storage of up to 10 months, this material has great potential for replacing and regenerating damaged stromal tissues.

Osteoinductive Moldable and Curable Bone Substitutes Based on Collagen, BMP-2 and Highly Porous Polylactide Granules, or a Mix of HAP/ß-TCP.

In dentistry, maxillofacial surgery, traumatology, and orthopedics, there is a need to use osteoplastic materials that have not only osteoinductive and osteoconductive properties but are also convenient for use. In the study, compositions based on collagen hydrogel were developed. Polylactide granules (PLA) or a traditional bone graft, a mixture of hydroxyapatite and ß-tricalcium phosphate (HAP/ß-TCP), were used for gel filling to improve mechanical osteoconductive properties of compositions. The mechanical tests showed that collagen hydrogels filled with 12 wt% highly porous PLA granules (elastic modulus 373 ± 55 kPa) or 35 wt% HAP/ß-TCP granules (elastic modulus 451 ± 32 kPa) had optimal manipulative properties. All composite components were cytocompatible. The cell's viability was above 90%, and the components' structure facilitated the cell's surface adhesion. The bone morphogenetic protein-2 (BMP-2) provided osteoinductive composition properties. It was impregnated directly into the collagen hydrogel with the addition of fibronectin or inside porous PLA granules. The implantation of a collagen hydrogel with BMP-2 and PLA granules into a critical-size calvarial defect in rats led to the formation of the most significant volume of bone tissue: 61 ± 15%. It was almost 2.5 times more than in the groups where a collagen-fibronectin hydrogel with a mixture of HAP/ß-TCP (25 ± 7%) or a fibronectin-free composition with porous PLA granules impregnated with BMP-2 (23 ± 8%) were used. Subcutaneous implantation of the compositions also showed their high biocompatibility and osteogenic potential in the absence of a bone environment. Thus, the collagen-fibronectin hydrogel with BMP-2 and PLA granules has optimal biocompatibility, osteogenic, and manipulative properties.

A new collagen scaffold for the improvement of corneal biomechanical properties in a rabbit model.

Although collagen based materials are widely used in corneal tissue engineering with promising results. The usage of such materials for the improvement of corneal biomechanical properties is still unclear. In this study, we aimed to investigate a new Viscoll collagen-based membrane for the improvement of corneal biomechanical characteristics.The right eyes of 15 Chinchilla rabbits were implanted with the membrane via an intrastromal pocket, with the contralateral intact eyes as controls. At 7, 30, 90, and 180 days post-surgery, the rabbits underwent anterior segment optical coherence topography, clinical examination, and slit-lamp microscopy. Additionally, the corneal samples also underwent histological examination followed by the assessment of the biomechanical characteristics of four treated and non-treated corneas at 30, 90, and 180 days, including keratometry at 180 days, post operation. Data are presented as means ± confidence intervals with a 95% confidence level. All the operated corneas retained their transparency throughout the study. Implantation approximately doubled the central corneal thickness. Corneas became stronger by approximately 87% between 1 and 6 months after surgery (maximum fracture load, 13.3 ± 0.8 and 24.9 ± 1.4 N, respectively), and their elasticity increased by approximately 27% over the same time frame (maximum slope of the elastic region of the stress-strain curve, 11.5 ± 0.2 and 14.6 ± 1.4 N/mm respectively). We have thus proposed a new method to increase corneal thickness and strengthen the corneal tissues while preserving their transparency and demonstrated its safety and efficacy in a rabbit model over 6 months. This may be a suitable alternative to the existing corneal collagen crosslinking procedure.

Collagen as Bioink for Bioprinting: A Comprehensive Review.

Biomaterials made using collagen are successfully used as a three-dimensional (3D) substrate for cell culture and considered to be promising scaffolds for creating artificial tissues. An important task that arises for engineering such materials is the simulation of physical and morphological properties of tissues, which must be restored or replaced. Modern additive technologies, including 3D bioprinting, can be applied to successfully solve this task. This review provides the latest evidence on advances of 3D bioprinting with collagen in the field of tissue engineering. It contains modern approaches for printing pure collagen bioinks consisting only of collagen and cells, as well as the obtained results from the use of pure collagen bioinks in different fields of tissue engineering.