The purpose of this study was to compare the use of autologous fibrin to human amniotic membrane (HAM) as a scaffold in cultivating autologous conjunctiva for transplantation in treatment of conjunctival defect. An experimental study was performed using 18 adult New Zealand white strain rabbits which were divided into 3 groups. Each group consists of 6 rabbits. The conjunctiva on the temporal site was excised to create a conjunctival epithelial defect. The excised area in the Group 1 was transplanted with autologous conjunctiva cultivated on autologous fibrin; Group 2 was transplanted with autologous conjunctiva cultivated on HAM and Group 3 was left bare. The rabbits were followed up at regular intervals until 6 weeks. The mean period of complete conjunctival epithelization was 11.50 ± 8.22 days for the autologous fibrin group, 15.33 ± 11.80 days for the HAM group and 25.33 ± 5.32 days in the bare sclera group. The epithelization rate for the autologous fibrin group was faster compared to the other two groups. However all the results were not statistically significant (p value >0.05). There were no postoperative complications noted during the follow up. Autologous fibrin is comparable to HAM as a scaffold for cultivation of conjunctiva in the treatment of conjunctival defect.
The proinflammatory cytokines, metalloproteinases family (MMPs), inflammatory mediators PGE2, COX-2 and NO are the most important group of compounds responsible for the loss of metabolic homeostasis of articular cartilage by promoting catabolic and destructive processes in the pathogenesis of osteoarthritis (OA). Stichopus chloronotus, a marine sea cucumber which is rich in n-3 PUFAs and phenolic compound, may exert a favorable influence on the course of the disease. The objective of this study was to investigate the regeneration and anti-inflammatory potential of S. chloronotus aqueous extract (SCAE) on human OA articular chondrocytes (HOC). Methods: The HOC isolated from knee joint cartilage removed during surgery were cultured with SCAE for 7 days. The effect of SCAE on anabolic and catabolic gene expression was verified by real-time PCR. Monolayer chondrocytes were stained with toluidine blue whereas sGAG, NO and PGE2 production in medium were analyzed by ELISA. Results: The HOC cultured in various SCAE have polygonal morphology maintaining their chondrocytes characteristic. SAE supplementation tested was found to be effective prochondrogenic, anti-inflammatory and anti-oxidative agents, as evidenced by upregulation of cartilage specific markers collagen type II, aggrecan core protein and sox-9 expression and downregulation of collagen type 1, IL-1, IL-6, IL-8, MMP-1, MMP-3, MMP-13, COX-2, iNOS and PAR-2 expression. The presence of SCAE in the culture was able to increase sGAG and reduce NO and PGE2 production significantly. Conclusions: These results suggested that SCAE demonstrated chondroprotective ability by suppressing catabolic activities, oxidative damage and effectively promoting chondrocytes growth.
Persistent epithelial defect is associated with delayed wound healing secondary to ineffective conventional treatment. Researches on natural products have produced promising results for better treatment options. Edible bird's nest (EBN) has been scientifically proven to be able to induce cell division and promote regeneration, but there has been no research done to see its effects on corneal cells. This study was done to investigate the effects of EBN in promoting corneal wound healing using a monolayer cell culture model. Corneal epithelial cells (CEC) were isolated from six New Zealand White rabbits and cultured until Passage 1. The cells were then divided into four different groups using two different media; the standard medium (CM) and basal medium (BM) with or without supplementation of 0.05% EBN extract. A corneal wound was created by using a 4 mm corneal trephine in each well-plate to mimic the corneal defect. The progress of wound healing was assessed through migration study. RT-PCR of genes associated with corneal epithelial wound healing (fibronectin, CD44 and Cytokeratin 3 (CK3)) were done to measure the expression level, and the protein expressions were further confirmed by immunocytochemistry. Cell migration study revealed the fastest wound closure occurred in culture using CM+0.05% EBN. Gene expression of fibronectin and CD44 were significantly reduced while CK3 expression was significantly increased in CEC cultured in CM+0.05% EBN. Only CD44 and CK3 proteins were detected in epithelial cells cultured in CM with 0.05% EBN while fibronectin was not detected. In conclusion, supplementation of EBN extract at 0.05% concentration in CEC culture media could promote cell migration, gene expressions and proteins associated with corneal wound healing.
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